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Information on EC 5.4.99.1 - methylaspartate mutase and Organism(s) Clostridium cochlearium and UniProt Accession P80078

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EC Tree
     5 Isomerases
         5.4 Intramolecular transferases
             5.4.99 Transferring other groups
                5.4.99.1 methylaspartate mutase
IUBMB Comments
Requires a cobamide coenzyme.
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This record set is specific for:
Clostridium cochlearium
UNIPROT: P80078
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Word Map
The taxonomic range for the selected organisms is: Clostridium cochlearium
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
glutamate mutase, adocbl-dependent glutamate mutase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Glutamate isomerase
-
-
-
-
Glutamate mutase
Glutamic acid isomerase
-
-
-
-
Glutamic acid mutase
-
-
-
-
Glutamic isomerase
-
-
-
-
Glutamic mutase
-
-
-
-
Methylaspartic acid mutase
-
-
-
-
Mutase, methylaspartate
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-threo-3-methylaspartate = L-glutamate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
isomerization
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-threo-3-Methylaspartate carboxy-aminomethylmutase
Requires a cobamide coenzyme.
CAS REGISTRY NUMBER
COMMENTARY hide
9032-97-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-Glu
L-threo-3-methylaspartate
show the reaction diagram
-
-
-
r
(2S,3S)-3-methylaspartate
L-glutamate
show the reaction diagram
-
-
-
-
r
(S)-2-hydroxyglutarate
(2S,3S)-3-methylmalate
show the reaction diagram
-
-
analysis of the energy profile for the various intermediate steps of reaction
-
?
Glu
?
show the reaction diagram
L-2-hydroxyglutarate
L-threo-3-methylmalate
show the reaction diagram
-
rate-limiting step is most likely the rearrangement of the 2-hydroxyglutaryl radical to the 3-methylmalyl radical
-
?
L-Glu
L-threo-3-methylaspartate
show the reaction diagram
-
-
-
r
L-Glu
threo-3-Methylaspartate
show the reaction diagram
L-glutamate
L-threo-3-methylaspartate
show the reaction diagram
L-threo-3-methylaspartate
L-glutamate
show the reaction diagram
additional information
?
-
-
the catalytic mechanism proceeds via a fragmentation/recombination sequence with intermediates stabilized by partial protonation/deprotonation
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
(2S,3S)-3-methylaspartate
L-glutamate
show the reaction diagram
-
-
-
-
r
Glu
?
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5'-deoxyadenosylcobalamin
-
dependent on
adenosylcobalamin
Cobalamin
coenzyme B12
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2S,3R)-3-Fluoroglutamate
-
-
(2S,4S)-4-Fluoroglutamate
-
-
(2S,4S)-4-Fluoroglutamic acid
-
-
(S)-2-thiolglutaric acid
-
high-level quantum chemical calculations of reaction intermediates
(S)-3-Methylitaconate
-
-
2-ketoglutaric acid
-
high-level quantum chemical calculations of reaction intermediates
2-Methyleneglutarate
-
binding of the 2-methyleneglutarate to glutamate mutase initiates homolysis of adensosylcobalamin, irreversible inhibition
2-Methyleneglutaric acid
-
-
2-oxoglutarate
-
inactivates, formation of the C-4 radical of 2-oxoglutarate is a facile process, but it does not undergo further reactions
2-thiolglutarate
-
competitive. 2-Thiolglutarate elicits cobalt-carbon bond homolysis and the formation of 5’-deoxyadenosine. 2-Thiolglutarate first forms a thiolglutaryl radical at C-4 that then undergoes fragmentation to produce acrylate and the sulfur-stabilized thioglycolyl radical which accumulates on the enzyme
Itaconic acid
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.2
L-2-hydroxyglutarate
-
pH 7.0
0.58 - 1.5
L-Glu
0.24 - 13
L-glutamate
0.14 - 9.5
L-threo-3-methylaspartate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.8
(2S,3S)-3-methylaspartate
-
genetically engineered enzyme with S subunit fused to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment
0.05
L-2-hydroxyglutarate
-
pH 7.0
5.8
L-Glu
-
genetically engineered enzyme with S subunit fused to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment
0.0181 - 6
L-glutamate
0.073 - 5.4
L-threo-3-methylaspartate
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
2-thiolglutarate
-
pH 7.0
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7
-
protein with the S subunit genetically fused to the C-terminus of the E-subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment
8 - 8.5
-
wild type enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 8.7
-
pH 6.5: about 50% of maximal activity, pH 8.7: about 80% of maximal activity
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
methylaspartate mutase S chain (subunit sigma)
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
residue Glu117 and the arginine claw have a strong influence, and also residues Glu 214, Lys 322, Gln 147, Glu 330, Lys 326, and Met 294 play a catalytic role. The arginine claw keeps the intermediates in place and is probably responsible for the enantioselectivity. Glu 171 temporarily accepts a proton from the glutamyl radical intermediate and donates it back at the end of the reaction
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
GMSS_CLOCO
137
0
14812
Swiss-Prot
-
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of inactive recombinant enzyme reconstituted with either cyanocobalamin or methylcobalamin, hanging-drop method
-
crystallization of component S, hanging drop method, polyethylene glycol 4000 as precipitant
-
recombinant enzyme
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E171A
-
turnover number for glutamate is reduced 27.6fold, KM-value is increased 1.1fold, Km-value for adenosylcobalamin is reduced 1.23fold
E171D
-
turnover number for glutamate is reduced 1.8fold, KM-value is reduced 1.54fold, Km-value for adenosylcobalamin is reduced 2.7fold
E171N
-
turnover number for glutamate is reduced 232fold, KM-value is increased by 1.8fold, Km-value for adenosylcobalamin is reduced 1.4fold
E171Q
-
turnover number for glutamate is reduced 53fold, KM-value is reduced 2.4fold, Km-value for adenosylcobalamin is reduced 2fold, mutant enzyme is independent of pH
R100K
R100M
-
no cob(II)alamin detected in UV-visible spectrum. Km-value for glutamate is reduced 276fold compared to wild-type enzyme, KM-value for glutamate is increased 13fold compared to wild-type enzyme
R100Y
-
no cob(II)alamin detected in UV-visible spectrum. Km-value for glutamate is reduced 322fold compared to wild-type enzyme, KM-value for glutamate is increased 17fold compared to wild-type enzyme
additional information
-
the S subunit is genetically fused to the C-terminus of the E subunit through an 11 amino acid 5-Gly linker segment. The affinity of adenosylcobalamine is unchanged, but the turnover-number and the Km-value for Glu in the conversion of L-Glu to (2S,3S)-3-methylaspartate are decreased by about a third
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purification of component E and component S
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
-
overexpression of polypeptide chains sigma and epsilon in Escherichia coli
-
polypeptide chains sigma and epsilon expressed in Escherichia coli
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Bothe, H.; Darley, D.J.; Albracht, S.P.; Gerfen, G.J.; Golding, B.T.; Buckel, W.
Identification of the 4-glutamyl radical as an intermediate in the carbon skeleton rearrangement catalyzed by coenzyme B12-dependent glutamate mutase from Clostridium cochlearium
Biochemistry
37
4105-4113
1998
Clostridium cochlearium
Manually annotated by BRENDA team
Switzer, R.L.
Glutamate mutase
B12 (Dolphin, D. ed. ) Wiley, New York
2
289-305
1982
Clostridium cochlearium, Clostridium saccharobutyricum, Clostridium sp., Acetoanaerobium sticklandii, Clostridium tetani, Clostridium tetanomorphum, no activity in Acidaminococcus fermentans, no activity in Clostridium microsporum, no activity in Fusobacterium fusiforme, no activity in Fusobacterium nucleatum, no activity in Micrococcus aerogenes, Cereibacter sphaeroides, Rhodospirillum rubrum, Clostridium sp. SB4
-
Manually annotated by BRENDA team
Barker, H.A.
Coenzyme B12-dependent mutases causing carbon chain rearrangements
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
6
509-537
1972
Cereibacter sphaeroides, Clostridium cochlearium, Clostridium tetanomorphum, Rhodospirillum rubrum
-
Manually annotated by BRENDA team
Chen, H.P.; Marsh, E.N.G.
Adenosylcobalamin-dependent glutamate mutase: examination of substrate and coenzyme binding in an engineered fusion protein possessing simplified subunit structure and kinetic properties
Biochemistry
36
14939-14945
1997
Clostridium cochlearium
Manually annotated by BRENDA team
Buckel, W.; Golding, B.T.
Glutamate and 2-methyleneglutarate mutase: from microbial curiosities to paradigms for coenzyme B12-dependent enzymes
Chem. Soc. Rev.
25
329-337
1996
Clostridium cochlearium, Clostridium tetanomorphum
-
Manually annotated by BRENDA team
Leutbecher, U.; Bcher, R.; Linder, D.; Buckel, W.
Glutamate mutase from Clostridium cochlearium
Eur. J. Biochem.
205
759-765
1992
Clostridium cochlearium
Manually annotated by BRENDA team
Reitzer, R.; Krasser, M.; Jogl, G.; Buckel, W.; Bothe, H.; Kratky, C.
Crystallization and preliminary X-ray analysis of recombinant glutamate mutase and of the isolated component S from Clostridium cochlearium
Acta Crystallogr. Sect. D
54
1039-1042
1998
Clostridium cochlearium
Manually annotated by BRENDA team
Zelder, O.; Beatrix, B.; Leutbecher, U.; Buckel, W.
Characterization of the coenzyme-B12-dependent glutamate mutase from Clostridium cochlearium produced in Escherichia coli
Eur. J. Biochem.
226
577-585
1994
Clostridium cochlearium
Manually annotated by BRENDA team
Roymoulik, I.; Moon, N.; Dunham, W.R.; Ballou, D.P.; Marsh, E.N.
Rearrangement of L-2-hydroxyglutarate to L-threo-3-methylmalate catalyzed by adenosylcobalamin-dependent glutamate mutase
Biochemistry
39
10340-10346
2000
Clostridium cochlearium
Manually annotated by BRENDA team
Huhta, M.S.; Ciceri, D.; Golding, B.T.; Marsh, E.N.
A novel reaction between adenosylcobalamin and 2-methyleneglutarate catalyzed by glutamate mutase
Biochemistry
41
3200-3206
2002
Clostridium cochlearium
Manually annotated by BRENDA team
Xia, L.; Ballou, D.P.; Marsh, E.N.
Role of arg100 in the active site of adenosylcobalamin-dependent glutamate mutase
Biochemistry
43
3238-3245
2004
Clostridium cochlearium
Manually annotated by BRENDA team
Madhavapeddi, P.; Marsh, E.N.
The role of the active site glutamate in the rearrangement of glutamate to 3-methylaspartate catalyzed by adenosylcobalamin-dependent glutamate mutase
Chem. Biol.
8
1143-1149
2001
Clostridium cochlearium
Manually annotated by BRENDA team
Gruber, K.; Kratky, C.
Coenzyme B(12) dependent glutamate mutase
Curr. Opin. Chem. Biol.
6
598-603
2002
Clostridium cochlearium
Manually annotated by BRENDA team
Hoffmann, B.; Konrat, R.; Bothe, H.; Buckel, W.; Krautler, B.
Structure and dynamics of the B12-binding subunit of glutamate mutase from Clostridium cochlearium
Eur. J. Biochem.
263
178-188
1999
Clostridium cochlearium
Manually annotated by BRENDA team
Chih, H.W.; Marsh, E.N.G.
Mechanism of glutamate mutase: identification and kinetic competence of acrylate and glycyl radical as intermediates in the rearrangement of glutamate to methylaspartate
J. Am. Chem. Soc.
122
10732-10733
2000
Clostridium cochlearium
-
Manually annotated by BRENDA team
Roymoulik, I.; Chen, H.P.; Marsh, E.N.
The reaction of the substrate analog 2-ketoglutarate with adenosylcobalamin-dependent glutamate mutase
J. Biol. Chem.
274
11619-11622
1999
Clostridium cochlearium
Manually annotated by BRENDA team
Reitzer, R.; Gruber, K.; Jogl, G.; Wagner, U.G.; Bothe, H.; Buckel, W.; Kratky, C.
Glutamate mutase from Clostridium cochlearium: the structure of a coenzyme B12-dependent enzyme provides new mechanistic insights
Structure
7
891-902
1999
Clostridium cochlearium
Manually annotated by BRENDA team
Cheng, M.C.; Marsh, E.N.
Pre-steady-state measurement of intrinsic secondary tritium isotope effects associated with the homolysis of adenosylcobalamin and the formation of 5'-deoxyadensosine in glutamate mutase
Biochemistry
43
2155-2158
2004
Clostridium cochlearium
Manually annotated by BRENDA team
Brooks, A.J.; Fox, C.C.; Marsh, E.N.; Vlasie, M.; Banerjee, R.; Brunold, T.C.
Electronic structure studies of the adenosylcobalamin cofactor in glutamate mutase
Biochemistry
44
15167-15181
2005
Clostridium cochlearium
Manually annotated by BRENDA team
Cheng, M.C.; Marsh, E.N.
Isotope effects for deuterium transfer between substrate and coenzyme in adenosylcobalamin-dependent glutamate mutase
Biochemistry
44
2686-2691
2005
Clostridium cochlearium
Manually annotated by BRENDA team
Sension, R.J.; Cole, A.G.; Harris, A.D.; Fox, C.C.; Woodbury, N.W.; Lin, S.; Marsh, E.N.G.
Photolysis and recombination of adenosylcobalamin bound to glutamate mutase
J. Am. Chem. Soc.
126
1598-1599
2004
Clostridium cochlearium
Manually annotated by BRENDA team
Sension, R.J.; Harris, D.A.; Stickrath, A.; Cole, A.G.; Fox, C.C.; Marsh, E.N.G.
Time-resolved measurements of the photolysis and recombination of adenosylcobalamin bound to glutamate mutase
J. Phys. Chem. B
109
18146-18152
2005
Clostridium cochlearium
Manually annotated by BRENDA team
Patwardhan, A.; Marsh, E.N.
Changes in the free energy profile of glutamate mutase imparted by the mutation of an active site arginine residue to lysine
Arch. Biochem. Biophys.
461
194-199
2007
Clostridium cochlearium
Manually annotated by BRENDA team
Yoon, M.; Patwardhan, A.; Qiao, C.; Mansoorabadi, S.O.; Menefee, A.L.; Reed, G.H.; Marsh, E.N.
Reaction of adenosylcobalamin-dependent glutamate mutase with 2-thiolglutarate
Biochemistry
45
11650-11657
2006
Clostridium cochlearium
Manually annotated by BRENDA team
Cheng, M.C.; Marsh, E.N.
Evidence for coupled motion and hydrogen tunneling of the reaction catalyzed by glutamate mutase
Biochemistry
46
883-889
2007
Clostridium cochlearium
Manually annotated by BRENDA team
Sandala, G.M.; Smith, D.M.; Marsh, E.N.; Radom, L.
Toward an improved understanding of the glutamate mutase system
J. Am. Chem. Soc.
129
1623-1633
2007
Clostridium cochlearium
Manually annotated by BRENDA team
Kozlowski, P.M.; Kamachi, T.; Kumar, M.; Nakayama, T.; Yoshizawa, K.
Theoretical analysis of the diradical nature of adenosylcobalamin cofactor-tyrosine complex in B12-dependent mutases: inspiring PCET-driven enzymatic catalysis
J. Phys. Chem. B
114
5928-5939
2010
Clostridium cochlearium (P80077), Clostridium cochlearium (P80078)
Manually annotated by BRENDA team
Rommel, J.B.; Kaestner, J.
The fragmentation-recombination mechanism of the enzyme glutamate mutase studied by QM/MM simulations
J. Am. Chem. Soc.
133
10195-10203
2011
Clostridium cochlearium
Manually annotated by BRENDA team
Rommel, J.B.; Liu, Y.; Werner, H.J.; Kaestner, J.
Role of tunneling in the enzyme glutamate mutase
J. Phys. Chem. B
116
13682-13689
2012
Clostridium cochlearium
Manually annotated by BRENDA team