Information on EC 3.1.1.61 - protein-glutamate methylesterase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
3.1.1.61
-
RECOMMENDED NAME
GeneOntology No.
protein-glutamate methylesterase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
mechanism
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
mechanism of regulation of enzyme activity by the N-terminal domain
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
the enzyme is a response regulator protein in the bacterial chemotaxis two-component signal transduction pathway
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
enzyme is not necessary for the aggregation of the chemoreceptor complex in vivo
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
the receptor pentapeptide-binding site is located at the juncture between the carboxyl-terminal end of alpha-5, the final piece of secondary structure in the regulatory domain, and the linker that connects the regulatory and catalytic domains
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
mechanism
Escherichia coli JM 109
-
-
protein L-glutamate O5-methyl ester + H2O = protein L-glutamate + methanol
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of carboxylic ester
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
protein-L-glutamate-O5-methyl-ester acylhydrolase
Hydrolyses the products of EC 2.1.1.77 (protein-L-isoaspartate(D-aspartate) O-methyltransferase), EC 2.1.1.78 (isoorientin 3'-O-methyltransferase), EC 2.1.1.80 (protein-glutamate O-methyltransferase) and EC 2.1.1.100 (protein-S-isoprenylcysteine O-methyltransferase).
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
CheB
P07330
-
CheB methylesterase
-
-
-
-
CheB methylesterase
-
-
chemotaxis receptor McpB
-
-
chemotaxis-specific methylesterase
-
-
-
-
esterase, methyl-accepting chemotaxis protein methyl-
-
-
-
-
esterase, protein methyl-
-
-
-
-
methyl-accepting chemotaxis protein
-
-
-
-
methylesterae CheB
-
-
methylesterase CheB
-
-
-
-
methylesterase CheB
-
-
PME
-
-
-
-
protein carboxyl methylesterase
-
-
-
-
protein methylesterase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
69552-31-4
-
93792-01-9
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain JM 109
-
-
Manually annotated by BRENDA team
Escherichia coli JM 109
strain JM 109
-
-
Manually annotated by BRENDA team
Salmonella enterica subsp. enterica serovar Typhimurium ST89pGK2
strain ST89pGK2
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
interaction mode between CheB and chemoreceptors, overview
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
CheB + H2O
?
show the reaction diagram
-
substrate Cheb is a chemoreceptor
-
-
?
methyl-accepting chemotaxis protein + H2O
chemotaxis protein + methanol
show the reaction diagram
-
-
-
-
?
methyl-accepting chemotaxis protein + H2O
chemotaxis protein + methanol
show the reaction diagram
-
-
-
-
?
methyl-accepting chemotaxis protein + H2O
chemotaxis protein + methanol
show the reaction diagram
Salmonella enterica subsp. enterica serovar Typhimurium ST89pGK2
-
-
-
-
?
methylated adrenocorticotropic hormone + H2O
adrenocorticotropic hormone
show the reaction diagram
-
-
-
-
?
methylated bovine gamma-globulin + H2O
bovine gamma-globulin + methanol
show the reaction diagram
-
-
-
-
?
methylated calmodulin + H2O
calmodulin + methanol
show the reaction diagram
-
-
-
-
?
methylated follicle-stimulating hormone + H2O
follicle-stimulating hormone + methanol
show the reaction diagram
-
-
-
-
?
methylated gelatin + H2O
gelatin + methanol
show the reaction diagram
-
-
-
-
?
methylated growth hormone + H2O
growth hormone + methanol
show the reaction diagram
-
-
-
-
?
methylated histones + H2O
histones + methanol
show the reaction diagram
-
-
-
-
?
methylated luteinizing hormone + H2O
luteinizing hormone + methanol
show the reaction diagram
-
-
-
-
?
methylated prolactin + H2O
prolactin + methanol
show the reaction diagram
-
-
-
-
?
ovalbumin methyl ester + H2O
ovalbumin + methanol
show the reaction diagram
-
-
-
-
?
methylated seminal-plasma proteins + H2O
seminal-plasma protein + methanol
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
in prokaryotes it plays a crucial role in bacterial chemotaxis
-
-
-
additional information
?
-
-
enzyme catalyses hydrolysis of receptor glutamine or methylglutamate side-chains to glutamic acid
-
-
-
additional information
?
-
-
responsible for the demethylation of the methyl-accepting chemotaxis proteins
-
-
-
additional information
?
-
-
demethylation of chemotactic membrane receptors
-
-
-
additional information
?
-
-
in eukaryotes the enzyme is possibly involved in leukocyte chemotaxis
-
-
-
additional information
?
-
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
additional information
?
-
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
additional information
?
-
-
cheB mutants show decreased clockwise and counterclockwise event durations and exhibits a significant increase in the flagellar switching frequencies
-
-
-
additional information
?
-
-
methylesterae CheB is involved in sensory adaption in bacterial chemotaxis
-
-
-
additional information
?
-
-
the regulation of activity of methylesterase CheB via phosphorylation is central to chemotactic adaption
-
-
-
additional information
?
-
-
the carboxyl-terminal linker is important for chemoreceptor function
-
-
-
additional information
?
-
-
surfaces with differing electrostatic properties may reflect CheB regions that mediate protein-protein interaction, computational docking, overview
-
-
-
additional information
?
-
Salmonella enterica subsp. enterica serovar Typhimurium ST89pGK2
-
responsible for the demethylation of the methyl-accepting chemotaxis proteins
-
-
-
additional information
?
-
Escherichia coli JM 109
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
CheB + H2O
?
show the reaction diagram
-
substrate Cheb is a chemoreceptor
-
-
?
additional information
?
-
-
in prokaryotes it plays a crucial role in bacterial chemotaxis
-
-
-
additional information
?
-
-
enzyme catalyses hydrolysis of receptor glutamine or methylglutamate side-chains to glutamic acid
-
-
-
additional information
?
-
-
responsible for the demethylation of the methyl-accepting chemotaxis proteins
-
-
-
additional information
?
-
-
demethylation of chemotactic membrane receptors
-
-
-
additional information
?
-
-
in eukaryotes the enzyme is possibly involved in leukocyte chemotaxis
-
-
-
additional information
?
-
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
additional information
?
-
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
additional information
?
-
-
cheB mutants show decreased clockwise and counterclockwise event durations and exhibits a significant increase in the flagellar switching frequencies
-
-
-
additional information
?
-
-
methylesterae CheB is involved in sensory adaption in bacterial chemotaxis
-
-
-
additional information
?
-
-
the regulation of activity of methylesterase CheB via phosphorylation is central to chemotactic adaption
-
-
-
additional information
?
-
Salmonella enterica subsp. enterica serovar Typhimurium ST89pGK2
-
responsible for the demethylation of the methyl-accepting chemotaxis proteins
-
-
-
additional information
?
-
Escherichia coli JM 109
-
enzyme catalyses the hydrolysis of glutamyl-methyl esters in bacterial chemoreceptor proteins
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
as effective as Mg2+ in activation
Mg2+
-
phosphorylation of intact enzyme requires Mg2+
Mg2+
-
divalent cation required; maximal activity with 1.1 mM Mg2+
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
D-Phe-Pro-Arg-H
-
-
Divalent ions
-
-
-
N-tert-Butoxycarbonyl-Gln-Leu-Lys-H
-
-
P2 domain of histidine kinase
-
inhibition is decreased upon phosphorylation
-
additional information
-
the unphosphorylated regulator domain partially inhibits methylesterase
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0016
-
deaminated calmodulin
-
pH 6.0, 37C
-
0.015
-
methyl-accepting chemotaxis protein
-
pH 7.0, 30C
-
0.0026
-
methylated adrenocorticotropic hormone
-
pH 6.0, 37C
-
0.063
-
methylated bovine gamma-globulin
-
pH 6.0, 37C
-
0.0009
-
methylated calmodulin
-
pH 6.0, 37C
-
0.0024
-
methylated follicle-stimulating hormone
-
pH 6.0, 37C
-
0.0013
-
methylated gelatin
-
pH 6.0, 37C
-
0.0174
-
methylated growth hormone
-
pH 6.0, 37C
-
0.0005
-
methylated histone
-
pH 6.0, 37C
0.0019
-
methylated luteinizing hormone
-
pH 6.0, 37C
-
1e-05
-
methylated methyl-accepting chemotaxis protein
-
pH 7.0, 22C
-
0.0072
-
methylated ovalbumin
-
pH 6.0, 37C
-
0.026
-
methylated prolactin
-
pH 7.0, 30C
-
0.0059
-
ovalbumin methyl ester
-
pH 6.0, 37C
-
0.0021
-
methylated seminal-plasma protein
-
pH 6.0, 37C
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2e-05
0.00056
several methylated protein substrates
-
-
-
0.004
-
methyl-accepting chemotaxis protein
-
-
-
additional information
-
additional information
-
-
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.00054
-
chymostatin
-
pH 6.0, 37C
3.5e-05
-
Leupeptin
-
pH 6.0, 37C
0.0025
-
P2 domain of histidine kinase
-
pH 7.0, 25C, in absence of phosphoramidate
-
0.021
-
P2 domain of histidine kinase
-
pH 7.0, 25C, in presence of phosphoramidate
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
37
-
50% of activity maximum at 20C and 37C, about 60% of activity maximum at 22C
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4.45
-
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
from caput epididymidis, level declines more than 20fold during maturation as spermatozoa acquire motility and capacity to fertilize
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
CheB localizes to a cell pole in presence of the chemoreceptor MCP, binding of the NL domain to the P2 domain targets methylesterase CheB to the polar signalling complex
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
31000
-
-
gel filtration
41000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Salmonella enterica subsp. enterica serovar Typhimurium ST89pGK2
-
x * 37000, SDS-PAGE
-
monomer
-
* 31000 SDS-PAGE
monomer
-
1 * 41000 SDS-PAGE
additional information
-
enzyme may be a serine hydrolase
additional information
-
the CheB protein is composed of at least 2 structurally distinct proportions: a C-terminal catalytic domain, and a N-terminal region which modulates esterase activity
additional information
-
the CheB protein is composed of at least 2 structurally distinct proportions: a C-terminal catalytic domain, and a N-terminal region which modulates esterase activity
additional information
-
the CheB protein is composed of at least 2 structurally distinct proportions: a C-terminal catalytic domain, and a N-terminal region which modulates esterase activity
additional information
-
the Thermotoga maritima CheB methylesterase domain has identical topology of a modified doubly-wound alpha/beta fold, but the CheB demethylation consensus sites of the chemoreceptors, the CheB substrate, are not uniquely conserved
additional information
Escherichia coli JM 109
-
enzyme may be a serine hydrolase
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
phosphoprotein
-
-
phosphoprotein
-
the regulation of activity of methylesterase CheB via phosphorylation is central to chemotactic adaption
phosphoprotein
-
phosphorylation by phosphoramidate requires Mg2+, phosphorylation of the regulatory domain at Asp-56 results in a reorganization of the domain interface, allowing exposure of the active site to the receptor substrate and simultaneously stimulating methylesterase activity
phosphoprotein
-
phosphorylation causes subtle changes in solvent accessibility at the interdomain interface of enzyme
phosphoprotein
-
enzyme is activated in vivo by phosphorylation of a single aspartate, Asp56, in its regulatory domain
phosphoprotein
-
In its unphosphorylated state, the regulatory domain inhibits enzyme activity of effector domain. Phosphorylation of the regulator domain leads to an enhancement of enzyme activity through a relief of inhibition and a stimulatory effect on catalysis.
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
crystal structure of the catalytic domain
-
the crystal structure model suggests a structural basis for phosphorylation-dependent activation of effector function
-
CheB methylesterase domain, X-ray diffraction structure determination and analysis at 2.2 A resolution
-
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
2
10
-
30 min, stable
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
stabilized by aqueous glycerol solution
-
maximal stability and low activity is observed in phosphate buffer at pH 7 with 50-100 mM added NaCl, while MES and low salt concentration afford minimum protection against inactivation
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70C, stable for at least 1 month
-
-20C, 10 mM sodium acetate buffer, pH 4.35, 50% glycerol, 0.1 mM DTT, purified enzyme stable
-
4C, 10% glycerol, 0.04% sodium azide, stable for at least several months
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
strain ST89pGK2, plasmid bearing strain overproducing esterase and transferase
-
recombinant enzyme
-
recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli
-
expression of the N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D54N
-
mutation of aspartate phosphorylation site causes no loss of activity, provided the enzyme is sufficiently induced
DELTA1-140
-
truncated enzyme form is active
S173C
-
no release of methanol, even when induced
C207A
-
mutant retains at least 50% of the wild-type enzyme activity
C207A/C309A
-
the double mutant retains at 70% of the wild-type enzyme activity
C309A
-
mutant retains at least 50% of the wild-type enzyme activity
S164C
-
mutant has less than 2% of the wild-type activity
C207A
Escherichia coli JM 109
-
mutant retains at least 50% of the wild-type enzyme activity
-
C207A/C309A
Escherichia coli JM 109
-
the double mutant retains at 70% of the wild-type enzyme activity
-
C309A
Escherichia coli JM 109
-
mutant retains at least 50% of the wild-type enzyme activity
-
S164C
Escherichia coli JM 109
-
mutant has less than 2% of the wild-type activity
-
A137T
-
3fold higher enzyme activity than wild-type enzyme in absence of phosphoramidate, in presence of phosphoramidate mutant and wild-type proteins show similar increase in enzyme activity
C207A
-
mutation has no effect on enzyme activity
C207S/C309S
-
wild-type and mutant enzyme show significantly greater activity than the unphosphorylated proteins
C309A
-
mutation causes a complete loss of enzyme activity
D56C/C207S/C309S
-
the half-life of mutant-SPO3 is 28 days, the unphosphorylated wild-type and mutant enzyme show the same enzyme activities
D56N
-
no activation of enzyme activity in the presence of phosphoramidate
G84V
-
mutation causes a complete loss of enzyme activity
H138R
-
2.5fold higher enzyme activity than wild-type enzyme in absence of phosphoramidate, in presence of phosphoramidate mutant and wild-type proteins show similar increase in enzyme activity
L153P
-
2fold higher enzyme activity than wild-type enzyme in absence of phosphoramidate, in presence of phosphoramidate mutant and wild-type proteins show similar increase in enzyme activity
additional information
-
constructs of the N- and C-terminal domains and investigation of interaction between the isolated regulatory and effector domains of enzyme
R255W
-
4fold higher enzyme activity than wild-type enzyme in absence of phosphoramidate, in presence of phosphoramidate mutant and wild-type proteins show similar increase in enzyme activity
additional information
-
constructs of the N- and C-terminal domains and investigation of interaction between the isolated regulatory and effector domains of enzyme