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Literature summary for 5.1.1.3 extracted from

  • May, M.; Mehboob, S.; Mulhearn, D.C.; Wang, Z.; Yu, H.; Thatcher, G.R.; Santarsiero, B.D.; Johnson, M.E.; Mesecar, A.D.
    Structural and functional analysis of two glutamate racemase isozymes from Bacillus anthracis and implications for inhibitor design (2007), J. Mol. Biol., 371, 1219-1237.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.1.3

Cloned(Commentary)

Cloned (Comment) Organism
cloned in Escherichia coli as a fusion protein with a 6xHis tag at the N-terminus Bacillus anthracis

Crystallization (Commentary)

Crystallization (Comment) Organism
crystallization trials are performed by the hanging-drop, vapor-diffusion method, X-ray structure analysis shows that RacE1 and RacE2 are both dimers with monomers arranged in a “tail-to-tail” orientation, RCSB Protein Data Bank: 2GZM Bacillus anthracis
crystallization trials are performed by the hanging-drop, vapor-diffusion method. X-ray structure analysis shows that RacE1 and RacE2 are both dimers with monomers arranged in a tail-to-tail orientation, RCSB Protein Data Bank: 2DWU Bacillus anthracis

Protein Variants

Protein Variants Comment Organism
V149A mutant V149A has about a 2fold higher kcat than wild-type RacE2 (67/sec versus 38/sec), and has a Km value for L-glutamate similar to that of RacE2 (4.6 mM versus 3.7 mM), in the reverse reaction, V149A has a higher kcat than RacE2 (4.9/sec versus 1.6/sec, respectively), and its Km value for D-glutamate is the same as that of RacE2 (0.2 mM) Bacillus anthracis

Inhibitors

Inhibitors Comment Organism Structure
(2R,4S)-2-amino-4-(2-naphthyl)methyl pentanedioic acid D-glutamate analog, good competitive inhibitor for RacE1; D-glutamate analog, only weak inhibitor of RacE2, but a potent competitive inhibitor for mutant V149A Bacillus anthracis
(2R,4S,E)-2-amino-4-(3-phenylprop-2-enyl)pentanedioic acid D-glutamate analog, good competitive inhibitor for RacE1; D-glutamate analog, only weak inhibitor of RacE2, but a potent competitive inhibitor for mutant V149A Bacillus anthracis
(4R,2S)-2-cinnamyl-4-amino-5-hydroxypentanoic acid D-glutamate analog, good competitive inhibitor for RacE1; D-glutamate analog, only weak inhibitor of RacE2, but a potent competitive inhibitor for mutant V149A Bacillus anthracis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information mutant V149A has a Km value for L-glutamate similar to that of RacE2 (4.6 mM versus 3.7 mM). In the reverse reaction, the Km of V149A for D-glutamate is the same as that of RacE2 (0.2 mM) Bacillus anthracis
0.2
-
 D-glutamate
-
 Bacillus anthracis
0.9
-
 D-glutamate
-
 Bacillus anthracis
3.7
-
 L-glutamate
-
 Bacillus anthracis
8
-
 L-glutamate
-
 Bacillus anthracis

Localization

Localization Comment Organism GeneOntology No. Textmining

Organism

Organism UniProt Comment Textmining
Bacillus anthracis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
protein extract is loaded on a HiTrap affinity column charged with Co2+, the His-tags are not removed as the enzyme activities with and without these tags are not significantly different Bacillus anthracis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-glutamate
-
 Bacillus anthracis D-glutamate
-
 r

Subunits

Subunits Comment Organism
dimer both enzymes RacE1 and Rac2 are dimers with monomers arranged in a tail-to-tail orientation which is determined by gel filtration Bacillus anthracis

Synonyms

Synonyms Comment Organism
glutamate racemase
-
 Bacillus anthracis
RacE1
-
 Bacillus anthracis
RacE2
-
 Bacillus anthracis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Bacillus anthracis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information mutant V149A has about a 2fold higher kcat than wild-type RacE2 (67/sec versus 38/sec). In the reverse reaction, V149A has a higher kcat than RacE2 (4.9/sec versus 1.6/sec, respectively) Bacillus anthracis
1.6
-
 D-glutamate
-
 Bacillus anthracis
3.9
-
 D-glutamate
-
 Bacillus anthracis
17.7
-
 L-glutamate
-
 Bacillus anthracis
38
-
 L-glutamate
-
 Bacillus anthracis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.2
-
 assay at Bacillus anthracis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0002
-
 (2R,4S,E)-2-amino-4-(3-phenylprop-2-enyl)pentanedioic acid potent competitive inhibitor for RacE2 mutant V149A Bacillus anthracis
0.0002
-
 (4R,2S)-2-cinnamyl-4-amino-5-hydroxypentanoic acid potent competitive inhibitor for RacE2 mutant V149A Bacillus anthracis
0.0003
-
 (2R,4S)-2-amino-4-(2-naphthyl)methyl pentanedioic acid potent competitive inhibitor for RacE2 mutant V149A Bacillus anthracis
0.0015
-
 (4R,2S)-2-cinnamyl-4-amino-5-hydroxypentanoic acid D-glutamate analog, good competitive inhibitor for RacE1 Bacillus anthracis
0.0035
-
 (2R,4S,E)-2-amino-4-(3-phenylprop-2-enyl)pentanedioic acid D-glutamate analog, good competitive inhibitor for RacE1 Bacillus anthracis
0.0046
-
 (2R,4S)-2-amino-4-(2-naphthyl)methyl pentanedioic acid D-glutamate analog, good competitive inhibitor for RacE1 Bacillus anthracis
0.064
-
 (2R,4S,E)-2-amino-4-(3-phenylprop-2-enyl)pentanedioic acid D-glutamate analog, only weak inhibitor of RacE2 Bacillus anthracis
0.073
-
 (4R,2S)-2-cinnamyl-4-amino-5-hydroxypentanoic acid D-glutamate analog, only weak inhibitor of RacE2 Bacillus anthracis
0.289
-
 (2R,4S)-2-amino-4-(2-naphthyl)methyl pentanedioic acid D-glutamate analog, only weak inhibitor of RacE2 Bacillus anthracis