Information on EC 2.3.1.142 - glycoprotein O-fatty-acyltransferase

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

EC NUMBER
COMMENTARY hide
2.3.1.142
-
RECOMMENDED NAME
GeneOntology No.
glycoprotein O-fatty-acyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
palmitoyl-CoA + mucus glycoprotein = CoA + O-palmitoylglycoprotein
show the reaction diagram
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-
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
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-
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SYSTEMATIC NAME
IUBMB Comments
fatty-acyl-CoA:mucus-glycoprotein fatty-acyltransferase
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CAS REGISTRY NUMBER
COMMENTARY hide
122191-29-1
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
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-
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Manually annotated by BRENDA team
glnA1 gene, Rv2220
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-
Manually annotated by BRENDA team
Sprague-Dawley
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-
Manually annotated by BRENDA team
diverse strains
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-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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in the absence of Erf4, the rate of hydrolysis of the active site palmitoyl thioester intermediate is increased, resulting in reduced palmitoyl transfer to a Ras2 substrate
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
glutathione-S-transferase + 7,8-diacetoxy-4-methylcoumarin
?
show the reaction diagram
glutathione-S-transferase + acetyl-CoA
?
show the reaction diagram
palmitoyl-CoA + adenylate cyclase toxin
CoA + palmitoyladenylate cyclase toxin
show the reaction diagram
palmitoyl-CoA + mucus glycoprotein
CoA + O-palmitoylglycoprotein
show the reaction diagram
RGS16 + palmitoyl-CoA
O-palmitoyl-RGS16 + CoA
show the reaction diagram
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RGS16 is the regulator of G protein signaling 16
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-
r
tubulin Galphai + palmitoyl-CoA
O-palmitoyl-tubulin Galphai + CoA
show the reaction diagram
-
-
-
-
r
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
glutathione-S-transferase + 7,8-diacetoxy-4-methylcoumarin
?
show the reaction diagram
-
-
-
-
?
palmitoyl-CoA + adenylate cyclase toxin
CoA + palmitoyladenylate cyclase toxin
show the reaction diagram
-
palmitoylation activates adenylate cyclase toxin
-
?
palmitoyl-CoA + mucus glycoprotein
CoA + O-palmitoylglycoprotein
show the reaction diagram
additional information
?
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METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaF
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activation
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
hydroxylamine
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octyl-beta-D-glucoside
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at concentrations below its critical micellar concentration
additional information
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L-methionine-S-sulfoximine, a potent inhibitor of GlnA1, and a structural analogue of glutamate, fails to inhibit TAase activity of rGlnA1
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ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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activation
dithiothreitol
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activation
Triton X-100
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activation
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
7,8-diacetoxy-4-methylcoumarin
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pH 6.5, 37C, recombinant GlnA1
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0.188
acetyl-CoA
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pH 6.5, 37C, recombinant GlnA1
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000934
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2.4
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liver plasma membrane preparation
228
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partially purified enzyme
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.4
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assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
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secreted protein
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Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
243000
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native PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
95
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5 min, partial inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol, 2 mM, stabilizes during purification
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dithiothreitol stabilizes during purification
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enzyme is inactivated by degrading or denaturaring agents, instable during final gel filtration purification step
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Erf4 and the C-terminal 58 amino acids of Erf2 are required for Erf2 stability
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glycerol, 25% v/v, stabilizes
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protease inhibitors stabilize
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SDS inactivates
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, partially purified enzyme, 0.15% v/v Triton X-100, 150 mM KCl, 20 mM Tris-HCl, pH 7.4, 1 mM EGTA, 0.01 mg/ml leupeptin, at least 3 months without loss of activity
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-80C, stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
gastric mucosa, solubilized with Triton X-100, affinity chromatography
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partial, 95fold from liver by preparation of plasma membrane fraction, solubilization with 0.15% Triton X-100, and sequential chromatography including affinity chromatography on a cerulin-based palmitoylation inhibitor ligand
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recombinant His-tagged Erf2 with fused FLAG-tagged Erf4 from strain RJY1827, solubilization by 0.8% dodecylmaltoside and urea, and ultracentrifugation, purification by nickel affinity chromatography and desalting gel filtration, recombinant FLAG-tagged Erf2 by the same method
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solubilized with Triton X-100 or Nonidet P-40
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cloning of rasp gene encoding a putative O-acyl transferase
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co-overexpression of His-tagged Erf2 with fused FLAG-tagged Erf4 in strain RJY1827, expression of FLAG-tagged Erf2 in strain RJY
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expression of wild-type CyaC acyltransferase and various mutants in Escherichia coli
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gene glnA1 or Rv2220, DNA and amino acid sequence determination and analysis, expression in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A140G
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changed selectivity of acyltransferase for the 2 acetylation sites of adenylate cyclase toxin, a mixture of bi and monoacetylated toxins modified either at both Lys860 and Lys983 or only at Lys860 is generated
A140V
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monoacetylation of adenylate cyclase toxin almost exclusively at Lys983
C67S
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no effect on adenylate cyclase toxin activation
H33D
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loss of adenlayte cyclase toxin acetylation activity
H33S
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loss of adenlayte cyclase toxin acetylation activity
R141K
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no effect on adenylate cyclase toxin activation
R141L
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no effect on adenylate cyclase toxin activation
S30R
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loss of adenlayte cyclase toxin acetylation activity
S30W
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loss of adenlayte cyclase toxin acetylation activity
S68T
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no effect on adenylate cyclase toxin activation
additional information
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construction of a fusion protein that places the 58 amino acids of Erf2 onto the C-terminus of Pfa4, another DHHC enzyme that resides in the endoplasmic reticulum membrane, the modification highly reduces the half-life of the engineered protein