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Literature summary for 2.7.7.43 extracted from

  • Horsfall, L.E.; Nelson, A.; Berry, A.
    Identification and characterization of important residues in the catalytic mechanism of CMP-Neu5Ac synthetase from Neisseria meningitidis (2010), FEBS J., 277, 2779-2790.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.7.43

Activating Compound

Activating Compound Comment Organism Structure
additional information no necessity of including a sulfhydryl reagent, such as dithiothreitol, in the assay reaction Neisseria meningitidis

Protein Variants

Protein Variants Comment Organism
D209A almost complete loss of activity. Residue involved in metal binding Neisseria meningitidis
D211A dramatic loss of activity. Residue involved in metal binding Neisseria meningitidis
F192A 8fold increase on Km value for CTP Neisseria meningitidis
F193A 3fold increase on Km value for CTP Neisseria meningitidis
K142A 10000fold reduction in kcat with an insignificant, fourfold, rise in Km for CTP Neisseria meningitidis
K142A dramatic loss of activity. Residue involved in metal binding Neisseria meningitidis
N175A 16fold increases in the Km value for CTP and 23fold for N-acetylneuraminate Neisseria meningitidis
Q104A 40fold inccrease in Km value for N-acetylneuraminate Neisseria meningitidis
Q104E dramatic loss of activity. Residue involved in metal binding Neisseria meningitidis
Q104L dramatic loss of activity. Residue involved in metal binding Neisseria meningitidis
Q104N dramatic loss of activity. Residue involved in metal binding Neisseria meningitidis
R173A 38fold increase in Km value for N-acetylneuraminate Neisseria meningitidis
Y179A 200fold decrease in kcat value Neisseria meningitidis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.017
-
 CTP wild-type, pH 7.5, 22°C Neisseria meningitidis
0.026
-
 CTP mutant D211A, pH 7.5, 22°C Neisseria meningitidis
0.031
-
 CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
0.04
-
 CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
0.048
-
 CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
0.052
-
 CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
0.066
-
 N-acetylneuraminate mutant D211A, pH 7.5, 22°C Neisseria meningitidis
0.068
-
 N-acetylneuraminate wild-type, pH 7.5, 22°C Neisseria meningitidis
0.11
-
 N-acetylneuraminate mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
0.15
-
 CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
0.27
-
 CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis
0.29
-
 N-acetylneuraminate mutant R173A, pH 7.5, 22°C Neisseria meningitidis
0.38
-
 N-acetylneuraminate mutant F192A, pH 7.5, 22°C Neisseria meningitidis
0.78
-
 N-acetylneuraminate mutant F193A, pH 7.5, 22°C Neisseria meningitidis
1.6
-
 N-acetylneuraminate mutant N175A, pH 7.5, 22°C Neisseria meningitidis
2.6
-
 N-acetylneuraminate mutant Q104A, pH 7.5, 22°C Neisseria meningitidis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ enzyme recruits two Mg2+ ions during the catalytic cycle Neisseria meningitidis

Organism

Organism UniProt Comment Textmining
Neisseria meningitidis P0A0Z8
-
-

Reaction

Reaction Comment Organism Reaction ID
CTP + N-acetylneuraminate = diphosphate + CMP-N-acylneuraminate typical ordered-sequential kinetic behaviour, enzyme does not bind N-acetylneuraminate in the absence of CTP Neisseria meningitidis
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CTP + N-acetylneuraminate
-
 Neisseria meningitidis diphosphate + CMP-N-acetylneuraminate
-
 ?
additional information no substrate: 2-keto-3-deoxy-D-glycero-D-galacto-nonulosonic acid, kcat/Km value is about 5000fold lower than that of N-acetylneuraminate Neisseria meningitidis ?
-
 ?

Synonyms

Synonyms Comment Organism
CMP-Neu5Ac synthetase
-
 Neisseria meningitidis
CNS
-
 Neisseria meningitidis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.001
-
 CTP mutant D211A, pH 7.5, 22°C Neisseria meningitidis
0.05
-
 CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.2
-
 CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
5
-
 CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
7
-
 CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
10.2
-
 CTP wild-type, pH 7.5, 22°C Neisseria meningitidis
20
-
 CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
25
-
 CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.0005
-
 N-acetylneuraminate mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
0.0009
-
 N-acetylneuraminate mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
0.0013
-
 CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
0.0063
-
 N-acetylneuraminate mutant F193A, pH 7.5, 22°C Neisseria meningitidis
0.052
-
 N-acetylneuraminate mutant F192A, pH 7.5, 22°C Neisseria meningitidis
0.072
-
 CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
0.09
-
 CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis
0.097
-
 CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
0.132
-
 CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
0.143
-
 CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
0.148
-
 N-acetylneuraminate wild-type, pH 7.5, 22°C Neisseria meningitidis
0.155
-
 N-acetylneuraminate mutant N175A, pH 7.5, 22°C Neisseria meningitidis
0.23
-
 N-acetylneuraminate mutant R173A, pH 7.5, 22°C Neisseria meningitidis
0.6
-
 CTP wild-type, pH 7.5, 22°C Neisseria meningitidis