Literature summary for 3.1.8.1 extracted from

Yeung, D.T.; Josse, D.; Nicholson, J.D.; Khanal, A.; McAndrew, C.W.; Bahnson, B.J.; Lenz, D.E.; Cerasoli, D.M.
Structure/function analyses of human serum paraoxonase (HuPON1) mutants designed from a DFPase-like homology model (2004), Biochim. Biophys. Acta, 1702, 67-77.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of PON1 mutant enzymes in Escherichia coli	Homo sapiens

Protein Variants

Protein Variants	Comment	Organism
C284D	no enzymic activity	Homo sapiens
C284D	site-directed mutagenesis, inactive mutant	Homo sapiens
D269E	no enzymic activity	Homo sapiens
D269E	site-directed mutagenesis, inactive mutant	Homo sapiens
D54N	no enzymic activity	Homo sapiens
D54N	site-directed mutagenesis, inactive mutant	Homo sapiens
E313A	slightly decreased activity	Homo sapiens
E313A	site-directed mutagenesis, the mutant enzyme shows altered kinetics compared to the wild-type enzyme	Homo sapiens
E314A	enzymic activity similar to wild-type	Homo sapiens
E314A	site-directed mutagenesis, the mutant enzyme shows slightly altered kinetics compared to the wild-type enzyme	Homo sapiens
F222A	site-directed mutagenesis, inactive mutant	Homo sapiens
F222D	no enzymic activity	Homo sapiens
F222D	site-directed mutagenesis, inactive mutant	Homo sapiens
F222Y	site-directed mutagenesis, the mutant enzyme shows altered kinetics compared to the wild-type enzyme	Homo sapiens
F222Y	increased activity with phenyl acetate, no activity with paraoxon	Homo sapiens
G11A	site-directed mutagenesis, the mutant enzyme shows slightly altered kinetics compared to the wild-type enzyme	Homo sapiens
G11A	slightly enhanced activity with phenyl acetate and paraoxon	Homo sapiens
G11C	site-directed mutagenesis, the mutant enzyme shows slightly altered kinetics compared to the wild-type enzyme	Homo sapiens
G11C	slightly enhanced activity with phenyl acetate and paraoxon	Homo sapiens
G11S	site-directed mutagenesis, the mutant enzyme shows slightly altered kinetics compared to the wild-type enzyme	Homo sapiens
G11S	slightly enhanced activity with phenyl acetate and paraoxon	Homo sapiens
H115W	site-directed mutagenesis, the mutant enzyme shows reduced activity and altered kinetics compared to the wild-type enzyme	Homo sapiens
H115W	no enzymic activity with phenyl acetate, increased activity with paraoxon	Homo sapiens
H115W/N133S	site-directed mutagenesis, the mutant enzyme shows reduced activity and altered kinetics compared to the wild-type enzyme	Homo sapiens
H115W/N133S	no enzymic activity with phenyl acetate, slightly decreased activity with paraoxon	Homo sapiens
H134W	no enzymic activity	Homo sapiens
H134W	site-directed mutagenesis, inactive mutant	Homo sapiens
H134Y	no enzymic activity	Homo sapiens
H134Y	site-directed mutagenesis, inactive mutant	Homo sapiens
H285D	no enzymic activity	Homo sapiens
H285D	site-directed mutagenesis, inactive mutant	Homo sapiens
H285Y	no enzymic activity	Homo sapiens
H285Y	site-directed mutagenesis, inactive mutant	Homo sapiens
L69F	no enzymic activity	Homo sapiens
L69F	site-directed mutagenesis, inactive mutant	Homo sapiens
N133S	enzymic activity similar to wild-type	Homo sapiens
N133S	site-directed mutagenesis, the mutant enzyme shows slightly altered kinetics compared to the wild-type enzyme	Homo sapiens
N168E	no enzymic activity	Homo sapiens
N168E	site-directed mutagenesis, inactive mutant	Homo sapiens
N224A	no enzymic activity	Homo sapiens
N224A	site-directed mutagenesis, inactive mutant	Homo sapiens
V304A	no enzymic activity	Homo sapiens
V304A	site-directed mutagenesis, inactive mutant	Homo sapiens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.14	-	paraoxon	mutant E313A, pH 10.5, 25°C	Homo sapiens
0.15	-	paraoxon	mutant H115W/N133S, pH 10.5, 25°C	Homo sapiens
0.16	-	paraoxon	wild-type, pH 10.5, 25°C	Homo sapiens
0.17	-	paraoxon	mutant N133S, pH 10.5, 25°C	Homo sapiens
0.18	-	paraoxon	mutant G11A and mutant G11S, pH 10.5, 25°C	Homo sapiens
0.19	-	paraoxon	mutant E314A and mutant G11C, pH 10.5, 25°C	Homo sapiens
0.42	-	paraoxon	mutant H115W, pH 10.5, 25°C	Homo sapiens

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	serum enzyme	Homo sapiens	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ca2+	dependent on, Ca2+ is required for activity and is involved in the catalytic mechanism via coordinating residues Asp54, Asn168, Asn224, and Asp269, structure, overview	Homo sapiens

Organism

Organism	UniProt	Comment	Textmining
Homo sapiens	-	-	-
Homo sapiens	P27169	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Homo sapiens
native PON1 from serum, overview, recombinant mutant enzymes from Escherichia coli, transient expression of mutant enzymes in HEK293 cells	Homo sapiens

Reaction

Reaction	Comment	Organism	Reaction ID
an aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol	structure-function analysis, catalytic mechanism involving the residue His285, residues Leu69, His134, Phe222, and Cys284 are involved in substrate binding, Ca2+ is required for activity and is involved in the catalytic mechanism via coordinating residues Asp54, Asn168, Asn224, and Asp269	Homo sapiens	a

Source Tissue

Source Tissue	Comment	Organism	Textmining
blood serum	-	Homo sapiens	-
serum	-	Homo sapiens	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
diethyl 4-nitrophenyl phosphate + H2O	-	Homo sapiens	4-nitrophenol + diethyl phosphate	-	?
paraoxon + H2O	-	Homo sapiens	4-nitrophenol + diethyl phosphate	-	?

Subunits

Subunits	Comment	Organism
More	secondary structure-function analysis with enzyme structure drawn from a DFPase-like homology model, cf. EC 3.1.8.2, overview	Homo sapiens

Synonyms

Synonyms	Comment	Organism
HuPON1	-	Homo sapiens
serum paraoxonase	-	Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Homo sapiens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Homo sapiens
10.5	-	assay at	Homo sapiens