Information on EC 5.4.99.4 - 2-Methyleneglutarate mutase

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The expected taxonomic range for this enzyme is: Eubacterium barkeri

EC NUMBER
COMMENTARY hide
5.4.99.4
-
RECOMMENDED NAME
GeneOntology No.
2-Methyleneglutarate mutase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
2-Methyleneglutarate = 2-methylene-3-methylsuccinate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
group transfer
-
-
intramolecular
-
isomerization
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Nicotinate and nicotinamide metabolism
-
-
nicotinate degradation III
-
-
SYSTEMATIC NAME
IUBMB Comments
2-methyleneglutarate carboxy-methylenemethylmutase
Requires a cobamide coenzyme.
CAS REGISTRY NUMBER
COMMENTARY hide
9059-10-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(R)-3-[methyl-H3]methylitaconate
2-methyleneglutarate
show the reaction diagram
-
-
-
r
(R,Z)-3-methyl[2'-H1]itaconate
2-methyleneglutarate
show the reaction diagram
-
-
-
r
(R,Z)-3-[methyl-H3]methyl[2'-H1]itaconate
2-methyleneglutarate
show the reaction diagram
-
-
-
r
2-methyleneglutarate
(R)-3-methylitaconate
show the reaction diagram
-
-
-
r
2-Methyleneglutarate
2-Methylene-3-methylsuccinate
show the reaction diagram
2-methylene[4-H2]glutarate
(R)-3-methylitaconate
show the reaction diagram
-
-
-
r
additional information
?
-
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
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
adenosylcobalamin
-
-
cob(II)alamin
-
-
cobamide
vitamin B12
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(Z)-glutaconate
-
mechanism-based inactivation. Inhibitor induces homolysis of the Co-C bond of coenzyme B12 to afford cob(II)alamin and the 5'-deoxyadenosyl radical. The 5'-deoxyadenosyl radical adds to the double bond of (Z)-glutaconate to afford a radical adduct and subsequent formation of aquocobalamin
1-Methyl-1,2-cis-cyclopropanedicarboxylate
1-Methyl-1,2-trans-cyclopropanedicarboxylate
1-methylcyclopropane-(1R,2R)-dicarboxylate
-
weak noncompetitive
-
buta-1,3-diene-2,3-dicarboxylate
-
mechanism-based inactivation. Inhibitor induces homolysis of the Co-C bond of coenzyme B12 to afford cob(II)alamin and the 5'-deoxyadenosyl radical. The 5'-deoxyadenosyl radical adds to a double bond of buta-1,3-diene-2,3-dicarboxylate to afford a relatively stable radical adduct in the active site of the enzyme
citrate
-
slight
glutaconate
iodoacetamide
Itaconate
L-Malate
-
competitive
Maleate
-
competitive
mesaconate
p-Chloromercuriphenylsulfonic acid
-
-
succinate
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
sulfhydryl groups
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.4
(R)-3-methylitaconate
-
pH 7.4
0.38
(R)-3-[methyl-H3]methylitaconate
-
pH 7.4
3.1 - 7.1
2-Methyleneglutarate
2.3
2-methylene[4-H2]glutarate
-
pH 7.4
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
18
(R)-3-methylitaconate
Eubacterium barkeri
-
pH 7.4
1.1
(R)-3-[methyl-H3]methylitaconate
Eubacterium barkeri
-
pH 7.4
17
(R,Z)-3-methyl[2'-H1]itaconate
Eubacterium barkeri
-
pH 7.4
1.4
(R,Z)-3-[methyl-H3]methyl[2'-H1]itaconate
Eubacterium barkeri
-
pH 7.4
30
2-Methyleneglutarate
Eubacterium barkeri
-
pH 7.4
2.3
2-methylene[4-H2]glutarate
Eubacterium barkeri
-
pH 7.4
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
13
1-methylcyclopropane-(1R,2R)-dicarboxylate
-
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.51
-
preparation which is heavily contaminated with methylitaconate isomerase
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 8
-
sharp decrease in activity below pH 7.5 and above pH 8
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
170000
-
sucrose density gradient centrifugation
290000
-
gel filtration
300000
-
gel filtration, sucrose density gradient centrifugation, nondenaturing PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
tetramer
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme rapidly decays, when stored at low protein concentrations, 1 mg/ml. Complete loss of activity after 3 days at 0C and 38% loss of activity after 6 days at -80C
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme containing cobalamin(II) is stable to oxygen and is an inactive form of the enzyme
-
3526
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80C, protein concentration 1 mg/ml, 38% loss of activity after 6 days
-
-80C, protein concentration 10 mg/ml, about 10% loss of activity after 6 months
-
0C, protein concentration 1 mg/ml, complete loss of activity after 3 days
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
large scale
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
overexpression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D483N
-
0.06%of wild-type activity
H464Q
-
35% of wild-type activity
H485Q
-
less than 0.03% of wild-type activity
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
denaturation with 8 M urea in the presence of 2 mM dithiothreitol followed by gel chromatography and renaturation affords an inactive enzyme which contains 40-50% of the initially bound cobalamin
-