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Information on EC 3.1.8.2 - diisopropyl-fluorophosphatase
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3.1.8.2 diisopropyl-fluorophosphataseIUBMB Comments
Acts on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorus compounds (including 'nerve gases'). Inhibited by chelating agents; requires divalent cations. Related to EC 3.1.8.1 aryldialkylphosphatase.
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Word Map
- 3.1.8.2
- dfpase
-
organophosphorus
- dfp
- degradation
- squid
- sarin
- paraoxon
-
loligo
- environmental protection
- phosphotriesterases
-
opaas
-
bioscavenger
- arylesterase
- vx
- cyclosarin
- medicine
- methylphosphonofluoridate
-
2.8.2.1
-
2.3.1.5
-
carbanilate
- diisopropylphosphorofluoridate
- arylsulfotransferase
-
six-bladed
-
bioplastics
- udp-glucosyltransferase
- analysis
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
DFPase, di-isopropylphosphorofluoridate fluorohydrolase, dialkylfluorophosphatase, diisopropyl fluorophosphatase, diisopropyl phosphorofluoridate hydrolase, diisopropylfluorophosphatase, diisopropylfluorophosphonate dehalogenase, diisopropylphosphofluoridase, EC 3.8.2.1, h-PON1, 
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SYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
DFPase
di-isopropylphosphorofluoridate fluorohydrolase
-
-
-
-
dialkylfluorophosphatase
-
-
-
-
diisopropyl fluorophosphatase
diisopropyl phosphorofluoridate hydrolase
-
-
-
-
diisopropylfluorophosphatase
diisopropylfluorophosphonate dehalogenase
-
-
-
-
diisopropylphosphofluoridase
-
-
-
-
EC 3.8.2.1
-
-
formerly
-
isopropylphosphorofluoridase
-
-
-
-
OPA anhydrase
-
-
-
-
OPA anhydrolase
-
-
-
-
OPAA
OPAA-2
-
-
-
-
opd
OPH
organophosphate acid anhydrase
-
-
-
-
organophosphate anhydrolase/prolidase
organophosphorus acid anhydrolase
organophosphorus hydrolase
senescence marker protein-30
SMP30
somanase
-
-
-
-
tabunase
-
-
-
-
additional information
DFPase
-
-
-
-
DFPase
-
diisopropylfluorophosphatase
-
-
-
-
OPAA
-
-
-
-
organophosphorus acid anhydrolase
-
-
-
-
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
the enzyme exhibits stereoselectivity toward the hydrolysis of chiral substrates with a preference for the S-P enantiomers
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
catalytic mechanism, H287 is required for catalytic activity
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
enzyme exhibits stereoselectivity toward the chiral phosphorus center of the p-nitrophenyl analogs of sarin and soman
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
squide-type enzyme: diisopropyl fluorophosphate hydrolysed more rapidly than soman, Mazur-type enzyme: soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
calcium coordinating residue D229, in concert with direct substrate activation by the metal ion, renders the phosphorus atom of the substrate susceptible for attack of water, through generation of a phosphoenzyme intermediate
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
analysis of the catalytic reaction mechanism of the enzyme. Proposed mechanism for phosphoenzyme intermediate formation involving Asp229 as the nucleophile. (i) Asp229 attacks the phosphorus center of DFP to form a pentavalent intermediate and (ii) the P-F bond dissociates to form a tetrahedral phosphoenzyme intermediate. Hydrolysis of the phosphoenzyme intermediate is not shown. And proposed mechanism for hydrolysis involving an activated water as the nucleophile. (i) Asp229 abstracts a proton from a water molecule either stepwise or in concert as (ii) water attacks the phosphorus center, (iii) Glu21 abstracts a proton either stepwise or in concert as water forms a bond with phosphorus, and (iv) the P-F bond dissociates
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
catalytic reaction mechanism involving residue Asp269, molecular dynamics and modelling, overview
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
molecular details of the catalytic mechanism of h-PON1
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
the catalytic mechanism of DFPase is investigated using the hybrid density functional theory method B3LYP with a large quantum chemical model of the active site abstracted from the X-ray crystal structure. For the first step, two different pathways are considered: (1) residue Asp229 as a nucleophile inline attacks on the phosphorus center and (2) an activated water molecule as the nucleophile attacks on the phosphorus center. Both the Asp229 and the activated water molecule are capable of proceeding nucleophilic attack on the substrate in the presence of Ca2+ ion with the associated barriers 14.8 and 6.0 kcal/mol, respectively. The latter is much easier to perform the nucleophile attack. From the phosphoenzyme intermediate with the hexa-coordinated Ca2+, the uncoordinated Glu21 functions as a general base activated an additional water molecule to attack the carbon center of Asp229 and make the phosphate release. Residues Asn120 and Asn175 promote the elimination of the fluoride via donating strong hydrogen bonds. Residue Asp229 plays a dual role during the hydrolysis reaction process, either as a nucleophile or as a general base to activate the water nucleophile. The role of the calcium ion is providing a necessary conformation of the active site, facilitating the nucleophile formation and substrate orientation
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of phosphoric triester
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-
-
-
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SYSTEMATIC NAME
IUBMB Comments
diisopropyl-fluorophosphate fluorohydrolase
Acts on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorus compounds (including 'nerve gases'). Inhibited by chelating agents; requires divalent cations. Related to EC 3.1.8.1 aryldialkylphosphatase.
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CAS REGISTRY NUMBER
COMMENTARY
9032-18-2
-
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(RS)-propan-2-yl methylphosphonofluoridate + H2O
4-carbamoylphenyl diethyl phosphate
i.e. sarin
-
-
?
(RS)-propan-2-yl methylphosphonofluoridate + H2O
isopropyl phosphate methylphosphonate + fluoride
(RS)-propan-2-yl methylphosphonofluoridate + H2O
propan-2-yl methylphosphonate + fluoride
i.e. sarin
-
-
?
1,2,2-trimethylpropyl methylphosphonofluoridoate + H2O
1,2,2-trimethylpropyl methylphosphonate + fluoride + H+
-
-
-
?
1-methylethyl methylphosphonofluoridoate + H2O
1-methylethyl methylphosphonate + fluoride + H+
-
-
-
?
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
3,3-dimethylbutan-2-yl methylphosphonate + fluoride
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
isopropyl methylphosphonate + fluoride
i.e. soman
-
-
?
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
Pinacolyl methylphosphonate + fluoride
i.e. soman
-
-
?
4-nitrophenylisopropyl phenylphosphinate + H2O
?
-
at 22% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
4-nitrophenylpropyl phenylphosphinate + H2O
?
-
at 24% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
cyclohexyl methylphosphonofluoridate + H2O
cyclohexyl methylphosphonate + fluoride
i.e. cyclosarin
-
-
?
cyclohexyl methylphosphonofluoridoate + H2O
cyclohexyl methylphosphonate + fluoride + H+
-
-
-
?
cyclohexylmethylphosphonofluoridate + H2O
cyclohexyl methylphosphonate + fluoride
i.e. cyclosarin
-
-
?
diethyl fluorophosphate + H2O
diethyl phosphate + fluoride
-
-
-
?
ethyl dimethylamidocyanophosphate + H2O
ethyl hydrogen dimethylphosphoramidate + HCN
-
-
-
?
ethyl dimethylphosphoramidocyanidate + H2O
ethyl hydrogen dimethylphosphoramidate + HCN
i.e. tabun
-
-
?
ethyl N,N-dimethylphosphoramidocyanidate + H2O
ethyl N,N-dimethylphosphoramide + cyanide
-
i.e. tabun
-
-
?
mipafox + H2O
?
-
i.e. N,N'-diisopropyl phosphorodiamidofluoridate, poor substrate
-
-
?
O-ethyl S-(2-diisopropylamino)ethyl methylphosphonothioate + H2O
?
-
an organophosphorous nerve agent
-
-
?
(RS)-propan-2-yl methylphosphonofluoridate + H2O
isopropyl phosphate methylphosphonate + fluoride
i.e. sarin
-
-
?
(RS)-propan-2-yl methylphosphonofluoridate + H2O
isopropyl phosphate methylphosphonate + fluoride
i.e. sarin
-
-
?
(S)-sarin + H2O
?
-
the calculated free energy barrier for hydrolysis of (S)-sarin by the mechanism for diiisopropyl fluorophosphate is highly unfavorable. Hydrolysis of (S)-sarin proceeds by a mechanism in which Asp229 could activate an intervening water molecule for nucleophilic attack on the substrate
-
?
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
3,3-dimethylbutan-2-yl methylphosphonate + fluoride
i.e. soman
-
-
?
3-[fluoro(methyl)phosphoryl]oxy-2,2-dimethylbutane + H2O
3,3-dimethylbutan-2-yl methylphosphonate + fluoride
i.e. soman
-
-
?
4-nitrophenyl-ethyl(phenyl)phosphinate + H2O
?
-
40% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
4-nitrophenyl-ethyl(phenyl)phosphinate + H2O
?
-
i.e. 4-nitrophenylethyl phenylphosphinate
-
-
?
4-nitrophenyl-ethyl(phenyl)phosphinate + H2O
?
-
at 46% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
4-nitrophenyl-ethyl(phenyl)phosphinate + H2O
?
-
Rangia cuneata (Mazur-type enzyme): better substrate than diisopropyl fluorophosphate
-
-
?
4-nitrophenyl-ethyl(phenyl)phosphinate + H2O
?
-
i.e. NPEPP
-
-
?
4-nitrophenyl-methyl(phenyl)phosphinate + H2O
?
-
at 49% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
4-nitrophenyl-methyl(phenyl)phosphinate + H2O
?
-
i.e. 4-nitrophenylmethyl phenylphosphinate
-
-
?
4-nitrophenyl-methyl(phenyl)phosphinate + H2O
?
-
at 28% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridine
-
-
-
?
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridine
i.e. CPO, a metabolite of chlorpyrifos that is used as a pesticide in agriculture industry
-
-
?
cyclohexyl methyl fluorophosphate + H2O
cyclohexyl methyl hydrogen phosphate + fluoride
-
(+)isomer, pH 7.2, 1 mM MnCl2
-
-
?
cyclohexyl methyl fluorophosphate + H2O
cyclohexyl methyl hydrogen phosphate + fluoride
-
(+)isomer, pH 7.2, 1 mM MnCl2
-
-
?
cyclohexyl methyl fluorophosphate + H2O
cyclohexyl methyl hydrogen phosphate + fluoride
-
(+)isomer, pH 7.2, 1 mM MnCl2
-
-
?
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
reaction of EC 3.1.8.1
-
-
?
diethyl-paraoxon + H2O
diethyl phosphate + 4-nitrophenol
reaction of EC 3.1.8.1, paraoxonase
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
soman hydrolysed more rapidly than diisopropyl fluorophosphate
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
highly toxic structural analogue of G-class type of nerve agents
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
spectrophotometric determination with phenol red
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
best substrate of squid enzyme
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
The mechanism for hydrolysis of diisopropyl fluorophosphate involves nucleophilic attack by Asp229 on phosphorus to form a pentavalent intermediate. P-F bond dissociation then yields a phosphoacyl enzyme intermediate in the rate-limiting step. A water molecule, coordinated to the catalytic Ca2+, donates a proton to Asp121 and then attacks the tetrahedral phosphoacyl intermediate to liberate the diisopropyl phosphate product
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
room temperature, pH 7.5
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
at about 30% of O-pinacolylmethylphosphonofluoridate hydrolysis
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
binding analysis, overview
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
at about 5-10% of soman hydrolysis
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
an analogue molecule of G-type warfare agents
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. O,O-diisopropyl phosphorofluoridate
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
i.e. DFP
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
-
SMP30 is important in diisopropyl phosphofluoridate detoxification
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
-
substrate of SMP30
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
SMP30 is important in diisopropyl phosphofluoridate detoxification
-
-
?
diisopropyl phosphorofluoridate + H2O
diisopropyl phosphate + fluoride
-
pH 8.0, 25°C, 1 mM MgCl2
-
-
?
diisopropyl phosphorofluoridate + H2O
diisopropyl phosphate + fluoride
pH 8.0, 25°C, 1 mM MgCl2
-
-
?
ethyl dimethylphosphoramidocyanidate + H2O
?
i.e. tabun
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphate + fluoride
-
i.e. cyclosarin
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphate + fluoride
-
i.e. cyclosarin
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphate + fluoride
-
i.e. cyclosarin
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphonate + fluoride
-
no reaction with the (-)isomer O-cyclohexyl methylphosphonofluoridate
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphonate + fluoride
-
no reaction with the (-)isomer O-cyclohexyl methylphosphonofluoridate
-
-
?
O-cyclohexyl methylphosphonofluoridate + H2O
O-cyclohexyl methylphosphonate + fluoride
-
no reaction with the (-)isomer O-cyclohexyl methylphosphonofluoridate
-
-
?
O-cyclohexylmethylphosphonofluoridate + H2O
?
-
at 114% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
O-isopropylmethylphosphonofluoridate + H2O
O-isopropylmethylphosphate + fluoride
-
i.e. sarin
-
-
?
O-isopropylmethylphosphonofluoridate + H2O
O-isopropylmethylphosphate + fluoride
i.e. sarin
-
-
?
O-isopropylmethylphosphonofluoridate + H2O
O-isopropylmethylphosphate + fluoride
-
i.e. sarin
-
-
?
O-isopropylmethylphosphonofluoridate + H2O
O-isopropylmethylphosphate + fluoride
i.e. sarin
-
-
?
O-isopropylmethylphosphonofluoridate + H2O
O-isopropylmethylphosphate + fluoride
-
i.e. sarin
-
-
?
O-pinacolyl methylphosphonofluoridate + H2O
O-pinacolyl methylphosphate + fluoride
-
i.e. soman
-
-
?
O-pinacolyl methylphosphonofluoridate + H2O
O-pinacolyl methylphosphate + fluoride
i.e. soman
-
-
?
O-pinacolyl methylphosphonofluoridate + H2O
O-pinacolyl methylphosphate + fluoride
i.e. soman
-
-
?
O-pinacolyl methylphosphonofluoridate + H2O
O-pinacolyl methylphosphate + fluoride
-
i.e. soman
-
-
?
p-nitrophenyl-soman + H2O
p-nitrophenol + soman
-
the enzyme is immobilized on a photoluminescent porous silicon platform
soman is O-(2,3,3)-trimethylpropyl methylphosphonofluoridate
-
r
p-nitrophenyl-soman + H2O
p-nitrophenol + soman
-
the enzyme is immobilized on a photoluminescent porous silicon platform
soman is O-(2,3,3)-trimethylpropyl methylphosphonofluoridate
-
r
paraoxon + H2O
diethylphosphate + 4-nitrophenol
-
i.e. diethyl-4-nitrophenyl phosphate, poor substrate
-
-
?
paraoxon + H2O
diethylphosphate + 4-nitrophenol
-
substrate are also 16 paraoxon analogs
-
-
?
paraoxon + H2O
diethylphosphate + 4-nitrophenol
-
i.e. diethyl-4-nitrophenyl phosphate, poor substrate
-
-
?
paraoxon + H2O
diethylphosphate + 4-nitrophenol
-
i.e. diethyl-4-nitrophenyl phosphate, poor substrate
-
-
?
phenyl acetate + H2O
phenol + acetate
pH 8.0, 25°C, 1 mM MgCl2
-
-
?
sarin + H2O
?
-
at 20% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
sarin + H2O
?
-
substrates are also p-nitrophenyl analogs of sarin, the enzyme exhibits a stereoselective preference for the R-P-enantiomers of sarin
-
-
?
sarin + H2O
?
-
at 38% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
sarin + H2O
?
-
at 10% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
soman + H2O
?
-
at 240% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
soman + H2O
?
-
substrates are also p-nitrophenyl analogs of soman, the enzyme exhibits a stereoselective preference for the R-P-enantiomers of soman
-
-
?
soman + H2O
?
-
i.e. O-1,2,2-trimethylpropylmethylphosphono fluoride
-
-
?
soman + H2O
?
-
10% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
soman + H2O
?
-
i.e. O-1,2,2-trimethylpropylmethylphosphofluoridate
-
-
?
tabun + H2O
?
-
i.e. N,N-dimethylethylphosphoramidocyanidate, at 124% the rate of diisopropyl fluorophosphate hydrolysis
-
-
?
additional information
?
-
-
not: 4-nitrophenylphosphonate, 4-nitrophenylphosphinorylcholine
-
-
?
additional information
?
-
-
4-nitrophenylacetate, bis(4-nitrophenyl)phosphate, tris(4-nitrophenyl)phosphate
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown, enzyme may, in nature, be used in peptide metabolism
-
-
?
additional information
?
-
-
the ability of OPAA to cleave both G-type nerve agents comes from the hydroxide ion in the metal center that facilitates nucleophilic attack on either the carbonyl oxygen of the phosphorus center of organophosphorus nerve agents. The enzyme also shows prolidase activity
-
-
?
additional information
?
-
-
the ability of OPAA to cleave both G-type nerve agents comes from the hydroxide ion in the metal center that facilitates nucleophilic attack on either the carbonyl oxygen of the phosphorus center of organophosphorus nerve agents. The enzyme also shows prolidase activity
-
-
?
additional information
?
-
-
the enzyme shows broad substrate specificity and is able to degrade organophosphorus compounds with P-O, P-CN, and P-F bonds and is the only enzyme known to cleave the P-S bond, which is characteristic of V-type nerve agents such as VX
-
-
?
additional information
?
-
-
OPH is able to degrade a broad list of some of the most toxic organophosphorous pesticides, such as paraoxon, and OP nerve agents including DFP, sarin, and soman
-
-
?
additional information
?
-
the enzyme also exhibits activity of EC 3.1.8.1, aryldialkylphosphatase, as well as of EC 3.1.8.2, diisopropyl-fluorophosphatase. Enzyme-substrate docking study, overview
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
additional information
?
-
-
not: methanesulfonyl fluoride, phenylmethanesulfonyl fluoride, monofluorophosphate, iso-octamethylpyrophosphoramide (i.e. ios-OMPA)
-
-
?
additional information
?
-
-
the enzyme shows broad substrate specificity and is able to degrade organophosphorus compounds with P-O, P-CN, and P-F bonds and is the only enzyme known to cleave the P-S bond, which is characteristic of V-type nerve agents such as VX
-
-
?
additional information
?
-
-
additional substrate: dipeptide Gly-Pro, prolidase activity of enzyme
-
-
?
additional information
?
-
human PON1 is a calcium-dependent promiscuous enzyme (phosphotriesterase, arylesterase and lactonase) with a wide range of substrates. Human PON1 is capable of hydrolyzing a broad range of organophosphorus compounds, including paraoxon, diisopropylfluorophosphate (DFP) and nerve agents such as sarin, soman and VX
-
-
?
additional information
?
-
the enzyme is also active with substrates of EC 3.1.1.81, quorum-quenching N-acyl-homoserine lactonase, and EC 3.1.8.1, paraoxonase, hydrolyzing organophosphorus compounds. Measurement of N-oxodecanoyl-DL-homoserine lactone (3O-C10AHL)-hydrolyzing activity (EC 3.1.1.81) of recombinant h-PON1 enzymes is determined by using a recombinant quorum-sensing reporter Escherichia coli strain
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
additional information
?
-
-
enzyme is involved in the synthesis of isoethionate from cysteine
-
-
?
additional information
?
-
-
diisopropyl fluorophosphatase acts on a variety of organophosphorus compounds
-
-
?
additional information
?
-
-
The enzyme also acts as Ca2+-dependent phosphotriesterase. The hydrolytic reaction catalyzed by DFPase leads to the formation of a phosphate or phosphonate and a fluoride ion, resulting in detoxification of the organophosphorus agent. Presence of a phosphoenzyme intermediate in the reaction mechanism, which involves direct nucleophilic attack by Asp229 on the substrate, but no metal-assisted water activation, overview
-
-
?
additional information
?
-
diisopropyl-fluorophosphatase (DFPase) from Loligo vulgaris is highly stable and robust biocatalyst for the hydrolysis of various chemical warfare agents such as sarin, soman, tabun, but no natural substrate for DFPase has been identified to date
-
-
?
additional information
?
-
the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs)
-
-
?
additional information
?
-
-
the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs)
-
-
?
additional information
?
-
DFPase from Loligo vulgaris effectively catalyzes the hydrolysis of the bond between phosphorus and the fluoride (or cyanide) leaving group
-
-
?
additional information
?
-
DFPase from Loligo vulgaris effectively catalyzes the hydrolysis of the bond between phosphorus and the fluoride (or cyanide) leaving group
-
-
?
additional information
?
-
the enzyme is not hydrolytically active against compounds with P-O or P-S leaving group bonds, except for some soman derivatives, and shows no efficient hydrolytic activity against lactones or esters
-
-
?
additional information
?
-
the squid phosphotriesterase diisopropyl fluorophosphatase (DFPase) from Loligo vulgaris shows relatively specific substrate preference, efficiently catalyzing the hydrolysis of diisopropyl fluorophosphate (DFP) and G-type nerve agents, including tabun (GA), sarin (GB), soman (GD), and cyclohexyl sarin (GF). The detoxification of the organophosphorous agent is achieved by the hydrolytic reaction producing a phosphate or phosphonate and a fluoride ion. The DFPase from squid central nervous system shows strong preference for the hydrolysis of P-F or P-CN bonds, which are absent in natural compounds
-
-
?
additional information
?
-
the squid phosphotriesterase diisopropyl fluorophosphatase (DFPase) from Loligo vulgaris shows relatively specific substrate preference, efficiently catalyzing the hydrolysis of diisopropyl fluorophosphate (DFP) and G-type nerve agents, including tabun (GA), sarin (GB), soman (GD), and cyclohexyl sarin (GF). The detoxification of the organophosphorous agent is achieved by the hydrolytic reaction producing a phosphate or phosphonate and a fluoride ion. The DFPase from squid central nervous system shows strong preference for the hydrolysis of P-F or P-CN bonds, which are absent in natural compounds
-
-
?
additional information
?
-
-
no activity of SMP30 with paraoxon, dihydrocoumarin, gamma-nonalactone, and delta-dodecanolactone
-
-
?
additional information
?
-
-
the ability of OPAA to cleave both G-type nerve agents comes from the hydroxide ion in the metal center that facilitates nucleophilic attack on either the carbonyl oxygen of the phosphorus center of organophosphorus nerve agents. The enzyme also shows prolidase activity
-
-
?
additional information
?
-
-
4-nitrophenylacetate, bis(4-nitrophenyl)phosphate, tris(4-nitrophenyl)phosphate
-
-
?
additional information
?
-
-
the ability of OPAA to cleave both G-type nerve agents comes from the hydroxide ion in the metal center that facilitates nucleophilic attack on either the carbonyl oxygen of the phosphorus center of organophosphorus nerve agents. The enzyme also shows prolidase activity
-
-
?
additional information
?
-
-
enzyme expression in liver decreases androgen-independently of aging
-
-
?
additional information
?
-
enzyme expression in liver decreases androgen-independently of aging
-
-
?
additional information
?
-
-
no activity of SMP30 with paraoxon, dihydrocoumarin, gamma-nonalactone, and delta-dodecanolactone
-
-
?
additional information
?
-
no activity of SMP30 with paraoxon, dihydrocoumarin, gamma-nonalactone, and delta-dodecanolactone
-
-
?
additional information
?
-
-
no activity with paraoxon, dihydrocoumarin, gamma-nonalactone and delta-dodecanolactone
-
-
?
additional information
?
-
no activity with paraoxon, dihydrocoumarin, gamma-nonalactone and delta-dodecanolactone
-
-
?
additional information
?
-
the substrates of enzyme OPH include organophosphate insecticides paraoxon, parathion, methylparathion, coumaphos, and diazinon, as well as potent nerve agents sarin, soman, and their analogue diisopropylfluorophosphate (DFP), cf. EC 3.1.8.1 and 3.1.8.2
-
-
?
additional information
?
-
-
no substrates are N-acetylvaline, N-acetylleucine, N-acetylmethionine or N-acetylalanine, sodium diphosphate, parathion, octamethylpyrophosphoramide (i.e. OMPA), triacetin, creatine phosphate, acetylcholine, butyrylcholine
-
-
?
additional information
?
-
-
Tetrahymena thermophila has 5 enzyme forms, some share characteristics of both the squid and the Mazur-type DFPase
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
chlorpyrifos oxon + H2O
diethyl phosphate + 3,5,6-trichloropyridine
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
-
-
-
-
?
diisopropyl fluorophosphate + H2O
diisopropyl phosphate + fluoride
an analogue molecule of G-type warfare agents
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
-
SMP30 is important in diisopropyl phosphofluoridate detoxification
-
-
?
diisopropyl phosphofluoridate + H2O
diisopropyl phosphate + fluoride
SMP30 is important in diisopropyl phosphofluoridate detoxification
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown, enzyme may, in nature, be used in peptide metabolism
-
-
?
additional information
?
-
-
the enzyme shows broad substrate specificity and is able to degrade organophosphorus compounds with P-O, P-CN, and P-F bonds and is the only enzyme known to cleave the P-S bond, which is characteristic of V-type nerve agents such as VX
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
additional information
?
-
-
the enzyme shows broad substrate specificity and is able to degrade organophosphorus compounds with P-O, P-CN, and P-F bonds and is the only enzyme known to cleave the P-S bond, which is characteristic of V-type nerve agents such as VX
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
additional information
?
-
-
enzyme is involved in the synthesis of isoethionate from cysteine
-
-
?
additional information
?
-
-
diisopropyl fluorophosphatase acts on a variety of organophosphorus compounds
-
-
?
additional information
?
-
diisopropyl-fluorophosphatase (DFPase) from Loligo vulgaris is highly stable and robust biocatalyst for the hydrolysis of various chemical warfare agents such as sarin, soman, tabun, but no natural substrate for DFPase has been identified to date
-
-
?
additional information
?
-
the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs)
-
-
?
additional information
?
-
-
the diisopropylfluorophosphatase (DFPase) from the ganglion and brain of Loligo vulgaris acts on P-F bonds present in some organophosphorus pesticides (OPs)
-
-
?
additional information
?
-
-
enzyme expression in liver decreases androgen-independently of aging
-
-
?
additional information
?
-
enzyme expression in liver decreases androgen-independently of aging
-
-
?
additional information
?
-
-
the natural substrates for enzyme are unknown
-
-
?
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Cd2+
Co2+
Mg2+
fully activates the hydrolysis of diisopropyl phosphofluoridate
Mn2+
Ni2+
Zn2+
additional information
Ca2+
required for activity, the wild type enzyme contains two Ca2+ ions
Ca2+
required, Ca2+ ion in active site, coordinated to the moiety of DFP, two glutamines (Asn120 and Asn175), two negatively charged residues (Asp229 and Glu21), and two water molecules, thus completes its hepta-coordinated. Therefore, the metal ion plays an essential role in the precise orientation and activation of the substrate. The active site solvent molecule coordinating the catalytic calcium ion is identified as a water molecule other than a hydroxide
Cd2+
slightly activates the hydrolysis of diisopropyl phosphofluoridate
Co2+
fully activates the hydrolysis of diisopropyl phosphofluoridate
Mn2+
the active site harbors the binuclear Mn2+ ions, three glycolate oxygens of the enzyme coordinate the two Mn2+ atoms. All the oxygens of the carboxylate side chains coordinate the Mn2+ ions in the more favorable syn configuration, binding structure analysis, detailed overview
Mn2+
fully activates the hydrolysis of diisopropyl phosphofluoridate
Zn2+
required two equivalents of Zn2+ per native enzyme molecule
additional information
-
activity is driven by a binuclear metal center in the C-terminal region
additional information
-
activity is driven by a binuclear metal center in the C-terminal region
additional information
-
activity is driven by a binuclear metal center in the C-terminal region
additional information
-
activity is driven by a binuclear metal center in the C-terminal region
additional information
-
activity is driven by a binuclear metal center in the C-terminal region
additional information
-
no effect by Ca2+ on SMP30, the enzyme does not bind to Ca2+ at all
additional information
no effect by Ca2+ on SMP30, the enzyme does not bind to Ca2+ at all
additional information
the binuclear metal center, comprised of either two equivalents of Zn2+ in the native enzyme or mixed metal ions Cd2+, Co2+, Mn2+, and Ni2+ in the metal-substituted catalysts, which generates the activated water molecules that initiate nucleophilic attack on the phosphorus atom of the substrate, resulting in the cleavage of phosphoester bond and the release of leaving group
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2-hydroxyquinoline
a specific reversible competitive inhibitor of h-PON1 that is known to bind in the active site of the enzyme and inhibit the hydrolytic activities of the enzyme
fluoride
product fluoride effectively inhibits organophosphorus hydrolase-catalyzed hydrolysis of diisopropylfluorophosphate (DFP) by a linear mixed inhibition, which is partially released by triethanolamine adding at initial or later reaction stage
N,N'-diisopropyldiamidofluorophosphate
i.e. mipafox or DDFP. the inhibitor's hydrolysis product, N,N'-diisopropyldiamidophosphate, is present in the cocrystal structure and bound by coordinating the binuclear metals and forming hydrogen bonds and nonpolar interactions with active site residues. An unusual common feature of the binding of the two ligands is the involvement of only one oxygen atom of the glycolate carboxylate and the product DDP tetrahedral phosphate in bridging the two Mn2+ ions, binding structure analysis, detailed overview
O,O-dicyclopentylphosphoroamidate
crystallization data, phosphoryl oxygen of inhibitor is directly coordinated to the catalytic calcium ion
EGTA
-
only slightly by Fe3+ or Ni2+, not by Cs2+, Cu2+, Mg2+, Ca2+ or Zn2+; reversible by Mn2+ or Co2+
EGTA
-
Mn2+ reverses and stimulates over pre-EGTA-treatment activity level, Co2+ partially restores; reversible by Mn2+ or Co2+; strong
Mipafox
-
i.e. N,N'-diisopropyl phosphorodiamidofluoridate, DFPase-1, DFPase-2 and DFPase-3; reversible
additional information
the activation of organophosphorus hydrolase by aminoalcohol buffers can be attributed to the reduction of fluoride inhibition, which would be beneficial to the hydrolase-based detoxification of organophosphofluoridate
-
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
triethanolamine
best activator, EDTA abolishes the triethanolamine-induced activation of enzyme OPH
CdCl2
1 mM, stimulates hydrolysis to a lesser extent, 38% activity, compared to MgCl2
CoCl2
1 mM, stimulates hydrolysis well, 102% activity, compared to MgCl2
additional information
-
an approach using recombinant enzyme encapsulated within sterically stabilized liposomes to enhance diisopropyl fluorophosphates degradation
-
additional information
presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 mg/l Co2+ ion increases the specific activity of the recombinant enzyme
-
additional information
-
presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 mg/l Co2+ ion increases the specific activity of the recombinant enzyme
-
additional information
organophosphorus hydrolase (OPH)-catalyzed hydrolysis of diisopropylfluorophosphate (DFP), an analogue molecule of G-type warfare agents, is activated by aminoalcohols. OPH reaction are remarkably increased in the buffers containing aminoalcohols with C2 between nitrogen and oxygen in their structures, including triethanolamine (TEA), diethanolamine, monoethanolamine, 1-amino-2-propanol, 2-amino-2-methyl-1-propanol, and triisopropanolamine. The activation of OPH by aminoalcohol buffers can be attributed to the reduction of fluoride inhibition, which would be beneficial to the hydrolase-based detoxification of organophosphofluoridate
-
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.39
Coumaphos
-
pH not specified in the publication, temperature not specified in the publication
0.45
diazinon
-
pH not specified in the publication, temperature not specified in the publication
0.46
fensulfothion
-
pH not specified in the publication, temperature not specified in the publication
0.08
methyl parathion
-
pH not specified in the publication, temperature not specified in the publication
0.68
O-cyclohexyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
0.43
O-ethyl S-(2-diisopropylamino)ethyl methylphosphonothioate
-
pH not specified in the publication, temperature not specified in the publication
0.24
parathion
-
pH not specified in the publication, temperature not specified in the publication
0.048
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
2.72
diisopropyl fluorophosphate
wild-type, pH 7.5, 25°C, in nitrogen atmosphere
2.92
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 1-amino-2-propanol
2.95
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 2-amino-2-methyl-1-propanol
2.99
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
2.99
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 1 mM NaF
3.01
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme
3.08
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethanol
3.1
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y
3.15
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM monoethanolamine
3.18
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methanol
3.2
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y
3.2
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y
3.35
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM diethanolamine
3.38
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM glycerol
3.57
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 2 mM NaF
3.63
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
3.87
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 200 mM thiodipropanol and 1 mM NaF
3.97
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodiglycol
4
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y, with 300 mM triethanolamine
4.06
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol and 1 mM NaF
4.1
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethanolamine
4.17
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine and 300 mM ethanol
4.2
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y, with 300 mM triethanolamine
4.21
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triisopropanolamine
4.23
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 100 mM thiodipropanol and 1 mM NaF
4.32
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine and 300 mM ethanol
4.32
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM triethanolamine
4.41
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine and 300 mM ethanol
4.44
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine
4.45
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 3 mM NaF
4.45
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine and 300 mM ethanol
4.5
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y, with 300 mM triethanolamine
4.52
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine
4.54
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 200 mM thiodipropanol and 2 mM NaF
4.56
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine
4.58
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol and 2 mM NaF
4.61
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 3-amino-1-propanol
4.67
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine
4.7
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
4.77
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol
4.87
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 100 mM thiodipropanol and 2 mM NaF
4.89
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol and 3 mM NaF
5.04
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 200 mM thiodipropanol and 3 mM NaF
5.58
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 100 mM thiodipropanol and 3 mM NaF
23.36
diisopropyl fluorophosphate
wild-type, presence of 1 mM O,O-dicyclopentylphosphoroamidate, pH 7.5, 25°C, in nitrogen atmosphere
65.45
diisopropyl fluorophosphate
wild-type, presence of 3 mM O,O-dicyclopentylphosphoroamidate, pH 7.5, 25°C, in nitrogen atmosphere
0.7
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
1.57
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
0.5
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
2.48
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
0.058
paraoxon
-
pH not specified in the publication, temperature not specified in the publication
1.27
paraoxon
-
pH not specified in the publication, temperature not specified in the publication
additional information
additional information
-
effects of pH, temperature and ionic strength on KM are studied
-
additional information
additional information
-
comparison of kinetic constants of paraoxon analogs
-
additional information
additional information
Michaelis-Menten kinetics
-
additional information
additional information
Michaelis-Menten kinetic analysis, substrate diisopropyl fluorophosphate in presence of aminoalcohol activators
-
additional information
additional information
reaction kinetics of wild-type and mutant enzymes, overview
-
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
610
Coumaphos
-
pH not specified in the publication, temperature not specified in the publication
176
diazinon
-
pH not specified in the publication, temperature not specified in the publication
67
fensulfothion
-
pH not specified in the publication, temperature not specified in the publication
189
methyl parathion
-
pH not specified in the publication, temperature not specified in the publication
652
O-cyclohexyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
0.3
O-ethyl S-(2-diisopropylamino)ethyl methylphosphonothioate
-
pH not specified in the publication, temperature not specified in the publication
630
parathion
-
pH not specified in the publication, temperature not specified in the publication
0.0115
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM glycerol
0.0117
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme
0.0123
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethanol
0.0123
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methanol
0.0142
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 3-amino-1-propanol
0.0148
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine
0.0151
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol
0.0155
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine and 300 mM ethanol
0.0163
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine
0.0178
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine and 300 mM ethanol
0.018
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine
0.0193
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine
0.0195
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 1-amino-2-propanol
0.0195
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine and 300 mM ethanol
0.0209
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM monoethanolamine
0.0211
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 2-amino-2-methyl-1-propanol
0.0216
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine and 300 mM ethanol
0.0295
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodiglycol
0.031
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y
0.0312
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM diethanolamine
0.0361
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triisopropanolamine
0.042
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethanolamine
0.0477
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM triethanolamine
0.048
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y
0.049
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y
0.142
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y, with 300 mM triethanolamine
0.182
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y, with 300 mM triethanolamine
0.217
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y, with 300 mM triethanolamine
9.69
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
20.35
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
208
diisopropyl fluorophosphate
-
pH 8, 35°C, effects of pH, temperature and ionic strength on kcat are studied
230
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
465
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
2091
diisopropyl fluorophosphate
wild-type, presence of 3 mM O,O-dicyclopentylphosphoroamidate, pH 7.5, 25°C, in nitrogen atmosphere
2107
diisopropyl fluorophosphate
wild-type, pH 7.5, 25°C, in nitrogen atmosphere
2111
diisopropyl fluorophosphate
wild-type, presence of 1 mM O,O-dicyclopentylphosphoroamidate, pH 7.5, 25°C, in nitrogen atmosphere
56
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
442
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
5
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
151
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
6.11
paraoxon
-
pH not specified in the publication, temperature not specified in the publication
3170
paraoxon
-
pH not specified in the publication, temperature not specified in the publication
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kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1600
Coumaphos
-
pH not specified in the publication, temperature not specified in the publication
390
diazinon
-
pH not specified in the publication, temperature not specified in the publication
150
fensulfothion
-
pH not specified in the publication, temperature not specified in the publication
2400
methyl parathion
-
pH not specified in the publication, temperature not specified in the publication
959
O-cyclohexyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
0.045
O-ethyl S-(2-diisopropylamino)ethyl methylphosphonothioate
-
pH not specified in the publication, temperature not specified in the publication
55000
paraoxon
-
pH not specified in the publication, temperature not specified in the publication
2600
parathion
-
pH not specified in the publication, temperature not specified in the publication
0.0031
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 3-amino-1-propanol
0.0032
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodipropanol
0.0033
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine
0.0034
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM glycerol
0.0035
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine
0.0035
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethylamine and 300 mM ethanol
0.0039
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme
0.0039
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methanol
0.004
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM diethylamine and 300 mM ethanol
0.004
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethanol
0.004
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine
0.0044
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine
0.0045
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM ethylamine and 300 mM ethanol
0.0052
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM methylamine and 300 mM ethanol
0.0066
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM monoethanolamine
0.0067
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 1-amino-2-propanol
0.0072
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM 2-amino-2-methyl-1-propanol
0.0074
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM thiodiglycol
0.0086
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triisopropanolamine
0.0093
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM diethanolamine
0.0098
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y
0.01
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethanolamine
0.011
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant wild-type enzyme, with 400 mM triethanolamine
0.0153
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y
0.016
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y
0.032
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant L140Y, with 300 mM triethanolamine
0.043
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y, with 300 mM triethanolamine
0.054
diisopropyl fluorophosphate
pH 8.0, 25°C, recombinant mutant F132Y/L140Y, with 300 mM triethanolamine
2.67
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1 wild-type enzyme
4.33
diisopropyl fluorophosphate
pH 10.5, 25°C, recombinant PON1-hFc fusion enzyme
77
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
9700
diisopropyl fluorophosphate
-
pH not specified in the publication, temperature not specified in the publication
80
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
282
O-isopropylmethylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
10
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
61
O-pinacolyl methylphosphonofluoridate
-
pH not specified in the publication, temperature not specified in the publication
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Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.125
O,O-dicyclopentylphosphoroamidate
wild-type, pH 7.5, 25°C, in nitrogen atmosphere
5.17
fluoride
pH 8.0, 25°C, recombinant wild-type enzyme, with 100 mM triethanolamine
5.45
fluoride
pH 8.0, 25°C, recombinant wild-type enzyme, with 200 mM triethanolamine
7.51
fluoride
pH 8.0, 25°C, recombinant wild-type enzyme, with 300 mM triethanolamine
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SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.527
0.998
with diisopropyl phosphorofluoridate as substrate, pH 8.0, 25°C, 1 mM MgCl2
1.512
110
substrate sarin, pH 8.0, temperature not specified in the publication
13.8
purified recombinant PON1 wild-type enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate
143
158
17.95
purified recombinant PON1-hFc fusion enzyme, pH 10.5, 25°C, substrate diisopropyl fluorophosphate
188
198
substrate cyclosarin, pH 8.0, temperature not specified in the publication
2.4
-
O-cyclohexyl methylphosphonofluoridate (-)isomer as substrate, pH 7.2, 1 mM MnCl2
2.7
-
O-cyclohexyl methylphosphonofluoridate (-)isomer as substrate, pH 7.2, 1 mM MnCl2
225
substrate diisopropyl fluorophosphate, pH 8.0, temperature not specified in the publication
29.2
-
O-cyclohexyl methylphosphonofluoridate (+)isomer as substrate, pH 7.2, 1 mM MnCl2
382.39
purified recombinant engineered His-tagged InaV-N-DFPase enzyme, substrate paraoxon, pH 7.2, 30°C
65.3
-
O-cyclohexyl methylphosphonofluoridate (+)isomer as substrate, pH 7.2, 1 mM MnCl2
82
substrate soman, pH 8.0, temperature not specified in the publication
additional information
additional information
-
pH dependence of the specific activity of the mutant H287N
additional information
a maximum number of 53560000 molecules of diisopropyl fluorophosphate is hydrolyzed per minute per whole cell in the presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 ppm Co2+ ion. 34.69 U/ml with substrate paraoxon, pH 7.2, 30°C
additional information
-
a maximum number of 53560000 molecules of diisopropyl fluorophosphate is hydrolyzed per minute per whole cell in the presence of glucose-mineral-salt (GMS) supplemented with tryptone and 100 ppm Co2+ ion. 34.69 U/ml with substrate paraoxon, pH 7.2, 30°C
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pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.5
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pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 9
-
the effect of pH 6.0-9.0 on kinetic constants kcat and KM is studied, KM shows no perceivable dependence on pH within this pH range, kcat increases with pH to a limiting value at pH 8.0
7.6 - 9
-
about half-maximal activity at pH 7.6 and about 60% of maximal activity at pH 9
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TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
25
30
additional information
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TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
bifunctional enzyme exhibiting additionally prolidase activity, E.C. 3.4.13.9
-
-
brenda
-
SwissProt
brenda
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SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
posterior, higher activity in glands of female than male squid
brenda
additional information
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LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
the enzyme circulates in the blood bound to high-density lipoprotein (HDL)
-
brenda
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GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the phosphotriesterase activity development between PON1, EC 3.1.8.1, and DFPase, EC 3.1.8.2, is investigated by using the hybrid density functional theory method B3LYP. Structure comparisons of evolutionarily related enzymes show that the mutation of Asn270 leads to the catalytic Ca2+ ion indirectly connecting the buried structural Ca2+ ion via hydrogen bonds in DFPase. It can reduce the plasticity of enzymatic structure, and possibly change the substrate preference from paraoxon (preferred substrate of PON1) to DFP (preferred substrate of DFPase), which implies an evolutionary transition from mono- to dinuclear catalytic centers, enzyme catalysis mechanism from an evolutionary perspective, overview
malfunction
insufficient organophosphate-hydrolyzing activity of native enzyme affirms the urgent need to develop improved variant(s) having enhanced organophosphate-hydrolyzing activity. Enzyme mutants show altered substrate specificity with increased activity against paraoxon and lactone substrates, overview
physiological function
additional information
physiological function
-
diisopropyl fluorophosphatase is a calcium-dependent phosphotriesterase that acts on a variety of highly toxic organophosphorus compounds, that act as inhibitors of acetylcholinesterase
physiological function
human paraoxonase 1 (h-PON1) is a serum enzyme that can hydrolyze a variety of substrates, including organophosphate (OP) compounds. PON1 can hydrolyze and inactivate a variety of organophosphate (OP) compounds, including certain OP pesticides and nerve agents (NAs). It is a potential candidate for the development of antidote against OP poisoning in humans. The enzyme possesses anti-inflammatory, anti-oxidative, anti-diabetic and quorum sensor-hydrolyzing activities, it is proposed that the lactonase activity of the enzyme is important for these defensive roles, cf. EC 3.1.1.81
physiological function
the enzyme efficiently catalyze the hydrolysis of the substrate diisopropyl fluorophosphate and a wide range of organophosphorus nerve agents, including soman, sarin, and tabun
physiological function
the squid phosphotriesterase diisopropyl fluorophosphatase (DFPase) from Loligo vulgaris shows relatively specific substrate preference, efficiently catalyzing the hydrolysis of diisopropyl fluorophosphate (DFP) and G-type nerve agents, including tabun (GA), sarin (GB), soman (GD), and cyclohexyl sarin (GF). The detoxification of the organophosphorous (OP) agent is achieved by the hydrolytic reaction producing a phosphate or phosphonate and a fluoride ion
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
additional information
the OPAA structure is composed of two domains, amino and carboxy domains, with the latter exhibiting a pita bread architecture and harboring the active site with the binuclear Mn2+ ions. The native enzyme structure reveals the presence of a well-defined nonproteinaceous density in the active site, which might be due to a bound glycolate, which is isosteric with a glycine product. All three glycolate oxygens coordinate the two Mn2+ atoms
additional information
h-PON1 is a polymorphic enzyme. Molecular docking analysis, homology modelling, overview
additional information
in detoxification of nerve gas compounds or pesticides in the human body, due to non-human origin of the enzyme, immunological reactions occur when it is injected into body. In order to using DFPase as in vivo detoxifying agent, some manipulations to augment of its efficiency and to decrease of immunogenic problems are needed. Modifications such as PEGylation is one of the possible solutions to conquer these problems
additional information
residue Asp229 plays a role in the coordination variation of calcium during the reaction. Quantum mechanical/molecular mechanical umbrella sampling simulations displays that the hydrolysis of diisopropyl fluorophosphate (DFP) and (S)-sarin processes by DFPase presents two different reaction pathways involving nucleophilic attack by Asp229 or an activated water on phosphorus. Modeling of active site and reaction mechanism with nucleophile Asp229 and coordinating Ca2+, detailed overview. Optimized geometries for the intermediates, transition state, and product for the hydrolysis step of DFPase
additional information
residue Asp229 plays a role in the coordination variation of calcium during the reaction. Quantum mechanical/molecular mechanical umbrella sampling simulations displays that the hydrolysis of diisopropyl fluorophosphate (DFP) and (S)-sarin processes by DFPase presents two different reaction pathways involving nucleophilic attack by Asp229 or an activated water on phosphorus. Modeling of active site and reaction mechanism with nucleophile Asp229 and coordinating Ca2+, detailed overview. Optimized geometries for the intermediates, transition state, and product for the hydrolysis step of DFPase
additional information
the metal-substituted catalysts generates the activated water molecules that initiate nucleophilic attack on the phosphorus atom of the substrate, resulting in the cleavage of phosphoester bond and the release of leaving group
additional information
the phosphotriesterase diisopropyl fluorophosphatase (DFPase) is a calcium-dependent beta-propeller protein. PON1, EC 3.1.8.1, and DFPase, EC 3.1.8.2, seem to employ similar catalytic mechanisms as phosphotriesterase, due to their structural similarities of active sites. The attacking nucleophile for phosphotriester hydrolysis is identified to be an activated water molecule, with the nucleophile attacking the phosphorus center. The E53Q and D269N mutants in PON1 both possess measurable lactonase and paraoxonase activity, and mutation studies combined with related molecular dynamics simulations suggest that the water activated by Glu53 and Asp269 is the most likely attacking nucleophile. Analysis of the rate-determining reaction step of the organophosphorus compound hydrolysis catalyzed both by DFPase and PON1. Structure-function relationship, overview. Active site structure of DFPase (PDB ID 2GVW) and substrate docking
additional information
the phosphotriesterase diisopropyl fluorophosphatase (DFPase) is a calcium-dependent beta-propeller protein. PON1, EC 3.1.8.1, and DFPase, EC 3.1.8.2, seem to employ similar catalytic mechanisms as phosphotriesterase, due to their structural similarities of active sites. The attacking nucleophile for phosphotriester hydrolysis is identified to be an activated water molecule, with the nucleophile attacking the phosphorus center. The E53Q and D269N mutants in PON1 both possess measurable lactonase and paraoxonase activity, and mutation studies combined with related molecular dynamics simulations suggest that the water activated by Glu53 and Asp269 is the most likely attacking nucleophile. Analysis of the rate-determining reaction step of the organophosphorus compound hydrolysis catalyzed both by DFPase and PON1. Structure-function relationship, overview. Active site structure of DFPase (PDB ID 2GVW) and substrate docking
additional information
-
organophosphorus hydrolase, OPH, contains a two-oxygen bridging mechanism in the active site suggesting subtle differences compared to organophosphorus acid anhydrolase, OPAA. OPAAs exhibit higher soman activities, whereas OPHs have higher activity against the organophosphorus pesticide paraoxon
-
additional information
-
the OPAA structure is composed of two domains, amino and carboxy domains, with the latter exhibiting a pita bread architecture and harboring the active site with the binuclear Mn2+ ions. The native enzyme structure reveals the presence of a well-defined nonproteinaceous density in the active site, which might be due to a bound glycolate, which is isosteric with a glycine product. All three glycolate oxygens coordinate the two Mn2+ atoms
-
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UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). DFPA_LOLVU
314
0
35080
Swiss-Prot
other Location (Reliability: 2)
A0A023JCX3_PHODC
680
0
76063
TrEMBL
other Location (Reliability: 2)
A0A023JCX7_PHODC
741
0
83095
TrEMBL
other Location (Reliability: 2)
I0ICV4_PHYMF
Phycisphaera mikurensis (strain NBRC 102666 / KCTC 22515 / FYK2301M01)
279
0
29817
TrEMBL
-
A0A023JCX6_PHODC
460
0
52211
TrEMBL
Secretory Pathway (Reliability: 4)
OPD_SPHSA
Sphingobium fuliginis (strain ATCC 27551)
365
1
39004
Swiss-Prot
-
PON1_HUMAN
355
0
39731
Swiss-Prot
Secretory Pathway (Reliability: 4)
RGN_RAT
299
0
33390
Swiss-Prot
other Location (Reliability: 2)
Q96XE7_SULTO
Sulfurisphaera tokodaii (strain DSM 16993 / JCM 10545 / NBRC 100140 / 7)
201
0
23415
TrEMBL
-
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PDB
SCOP
CATH
UNIPROT
ORGANISM
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MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20000 - 30000
-
three enzyme types of different MW: 20000-30000, 45000-50000 and 70000-100000
34000
35000
45000 - 50000
-
three enzyme types of different MW: 20000-30000, 45000-50000 and 70000-100000
58000
x * 58000, the enzyme OPAA structure is composed of two domains, a small N-terminal domain or N-domain, residue 1 to 160, and a large C-terminal domain or C-domain
70000 - 100000
-
three enzyme types of different MW: 20000-30000, 45000-50000 and 70000-100000
35000
-
2 * 35000, OPH from displays the TIM barrel fold in the active site loaded with Zn2+, overview
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SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
homodimer
-
2 * 35000, OPH from displays the TIM barrel fold in the active site loaded with Zn2+, overview
monomer
tetramer
additional information
?
x * 58000, the enzyme OPAA structure is composed of two domains, a small N-terminal domain or N-domain, residue 1 to 160, and a large C-terminal domain or C-domain
?
-
x * 58000, the enzyme OPAA structure is composed of two domains, a small N-terminal domain or N-domain, residue 1 to 160, and a large C-terminal domain or C-domain
-
?
x * 56847, wild-type enzyme, sequence calculation, x * 55000, recombinant wild-type enzyme, SDS-PAGE, x * 77000, recombinant engineered His-tagged InaV-N-DFPase enzyme, SDS-PAGE
tetramer
-
active enzyme, OPAA/prolidase from Alteromonas sp. JD6.5 displays the pita bread fold in the C-terminal region which houses the active metal site loaded with Mn2+, overview
tetramer
-
active enzyme, OPAA/prolidase from Alteromonas sp. JD6.5 displays the pita bread fold in the C-terminal region which houses the active metal site loaded with Mn2+, overview
-
additional information
features observed in the structure of PON1, i.e. the six-bladed beta-propeller scaffold, the three alpha-helices at the top of the propeller and the putative calcium-binding residues, are well conserved in the modelled structures
additional information
the molecular conformation of DFPase is similar to other phosphotriesterases such as paraoxonase 1 (PON1, EC 3.1.8.1)
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POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
side-chain modification
covalent attachment of polyethylene glycol (PEG) chains to enzymes is a prominent solution to overcome these issues. PEGylation enlarges the hydrodynamic radius of proteins and shields its surface. PEG modification increases the stability against proteases, reduces immunogenicity, and delays renal excretion significantly leading to prolonged half-life, reduced side effects, and increased pharmacological efficiency
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified OPAA, in acetate buffer equilibrated in a reservoir solution containing a much greater concentration of acetate, 270 mM ammonium acetate and 60 mM sodium acetate, pH 4.6, X-ray structure determination and analysis at 2.7 A resolution for the enzyme with bound inhibitor N,N'-diisopropyldiamidophosphate, and at 2.3 A resolution for the native enzyme
at 2.2 A resolution, after exchange of available H atoms by using D2O. Collection of time-of-flight wavelength-resolved Laue images
-
by vapour diffusion using a protein solution with 2 mM protein in 10 mM Tris, pH 7.5, 2 mM CaCl2, against a precipitation solution containing 11% PEG 4000 in 0.1 M MES, pH 6.5, at room temperature, X-ray diffraction structure determination and analysis at 2.2 A resolution. Comparison with structures of enzyme mutants, overview
-
homology modeling and docking of substrates. The two identical hydrophobic isopropyl groups in diisopropyl fluorophosphate bind to two different sub-pockets in the binding-site. Sub-pocket 1 is formed by residues P36, E37, I72, A74 and M90 while sub-pocket 2 is formed by F173, N175, T195, and W244
hydrodynamic model calculations based on the DFPase crystal structure from the native state enzyme structure in solution by use of different scattering methods, i.e. small-angle neutron scattering, overview
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in complex with inhibitor O,O-dicyclopentylphosphoroamidate. Phosphoryl oxygen of inhibitor is directly coordinated to the catalytic calcium ion
mutant enzymes are crystallized at room temperature by hanging drop vapor diffusion method, using 0.1 M Tris buffer pH 8.5, 2% tacsimate, 16% (w/v) PEG 3350 for mutant E37A/Y144A/R146A/T195M or 0.2 M KCl, 0.05 M HEPES buffer pH 7.5, 35% (w/v) pentaerythriol propoxylate for mutant E37D/Y144A/R16A/T195M
mutant enzymes N120D/N175D/D229N, E21QN120D/N175D/D229N, and D121E are crystallized by hanging drop vapor diffusion method, using 0.1 M MES buffer pH 6.5, 14-20% (w/v) PEG 3350
quantum mechanical/molecular mechanical umbrella sampling simulations. The mechanism for hydrolysis of diisopropyl fluorophosphate involves nucleophilic attack by Asp229 on phosphorus to form a pentavalent intermediate. P-F bond dissociation then yields a phosphoacyl enzyme intermediate in the rate-limiting step. A water molecule, coordinated to the catalytic Ca2+, donates a proton to Asp121 and then attacks the tetrahedral phosphoacyl intermediate to liberate the diisopropyl phosphate product. The calculated free energy barrier for hydrolysis of (S)-sarin by the same mechanism is highly unfavorable. Hydrolysis of (S)-sarin proceeds by a mechanism in which Asp229 could activate an intervening water molecule for nucleophilic attack on the substrate
structural characterization of a squid-type enzyme, the overall structure of this protein represents a six-fold beta propeller with two calcium ions bound in a central water-filled tunnel
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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H254R
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the active site mutation results in increased activity with soman and VX
H275L
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the active site mutation results in increased activity with soman and VX
H275V
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the active site mutation results in increased activity with soman and VX
H115W/R192K
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/D94H/S211T
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/L130F
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/M127I/D263H
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
H115W/R192K/A137T/S81R/P165A
site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type
L55M
natural polymorphism, the polymorphism at the 55th position of h-PON1 does not affect the catalytic properties of the enzyme
R192E
natural polymorphism, polymorphism at position 192 plays an important role in determining the substrate specificity and catalytic efficiency of the enzyme
Asp229/Asn120
site-directed mutagenesis, the mutant shows impaired catalytic activity or decreased substrate binding affinity
Asp229/Asn175
site-directed mutagenesis, the mutant shows impaired catalytic activity or decreased substrate binding affinity
D121E
the mutant displays 87% activity compared to the wild type enzyme
D229N/N120D
D229N/N175D
catalytically inactive, no change in the calcium coordinating environment
E21Q/N120D/N175D/D229N
the mutations lead to a loss of calcium binding and enzymatic activity
E37A/Y144A/R146A/T195M
the mutant shows increased turnover number and kcat/Km for diisopropyl fluorophosphate compared to the wild type enzyme
E37D/Y144A/R16A/T195M
the mutant shows increased turnover number and kcat/Km for diisopropyl fluorophosphate compared to the wild type enzyme
Glu21/Asn120
site-directed mutagenesis, the mutant shows impaired catalytic activity or decreased substrate binding affinity
Glu21/Asn175
site-directed mutagenesis, the mutant shows impaired catalytic activity or decreased substrate binding affinity
H181N
H274N
H287N
N120D/N175D/D229N
the mutations lead to a loss of calcium binding and enzymatic activity
F132Y
site-directed mutagenesis, the mutant exhibits a 3fold increased activity with diisopropyl fluorophosphate compared to wild-type
F132Y/ L140Y
site-directed mutagenesis, the mutant exhibits a several folds increased activity with diisopropyl fluorophosphate compared to wild-type
L140Y
site-directed mutagenesis, the mutant exhibits a 5fold increased activity with diisopropyl fluorophosphate compared to wild-type
additional information
D229N/N120D
catalytically inactive, no change in the calcium coordinating environment
H274N
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slight loss of activity in comparison to wild-type enzyme using pH Stat measurements, no alteration is observed with fluoride assay
additional information
a PTE mutant dubbed C23 is engineered, exhibiting reversed stereoselectivity and high catalytic efficiency (kcat/KM) for the hydrolysis of the toxic enantiomers of nerve agents VX, CVX, and VR. The other mutants A53, IV-A1, IV-H3, B141, and RD1-G83 are less effective, activities and mutant-substrate docking, overview. The only exception is IV-A1 with amiton. Most variants are highly efficient when hydrolyzing N,N-diisopropyl substrates and are 2fold less efficient with N,N-diethyl substrates and 5fold less efficient with N,N-dimethyl substrates
additional information
h-PON1 is a polymorphic enzyme. A random mutagenesis approach is used to increase the organophosphate (OP)-hydrolyzing activity of recombinant enzyme h-PON1. The mutants not only show a 10-340fold increased OP-hydrolyzing activity against different OP substrates but also exhibit differential lactonase and arylesterase activities, molecular docking studies, overview. Random mutagenesis using Escherichia coli XL-1 Red mutator strain. All mutations result in a considerable decrease in the delta-valerolactone-hydrolyzing activity of the enzyme
additional information
the catalytic efficiency of the recombinant human paraoxonase 1 fused to human immunoglobulin Fc domain, i.e. recombinant PON1-hFc, towards diisopropyl fluorophosphate (DFP) and paraoxon hydrolysis is 1.63 and 1.24fold higher, respectively, than the recombinant human wild-type PON1
additional information
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chemical modification of Tyr, Cys, Arg, Lys, Glu and Asp, this residues are not critical for catalysis
additional information
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preparation of bicontinuous microemulsions made of sugar surfactants as host systems for the DFPase. The microemulsion remains stable in the presence of the enzyme, scattering experiments. DFPase still has high activity in this complex reaction medium, overview
additional information
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reengineering of DFPase through rational design to bind and productively orient the more toxic SP stereoisomers of the nerve agents sarin and cyclosarin, creating a modified enzyme with enhanced overall activity and significantly increased detoxification properties
additional information
intracellular production of organophosphorus pesticides (OPs)-degrading enzymes or the use of native bacteria and fungi leads to a low degradation rate of OPs due to a mass transfer issue which reduces the overall catalytic efficiency. To overcome this challenge, DFPase is expressed on the surface of Escherichia coli for the first time by employing the N-terminal domain of the ice nucleation protein (InaV-N) as an anchoring motif. The recombinant DFPase is successfully located on the outer membrane and shows a significant ability for the biodegradation of diisopropylfluorophosphate (DFP) with a specific activity of 500 U/mg of wet cell weight. The recombinant cells can also degrade chlorpyrifos. No enzyme activity is measured by the fluoride ion-selective electrode in the control sample, inner membrane fraction, or cytoplasm fraction of pET-28a-InaV-N-DFPase cells. High potential of the InaV-N anchoring domain to produce an engineered bacterium that can be used in the bioremediation of pesticide-contaminated environments
additional information