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Literature summary 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

Activating Compound

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

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.43 0.017
-
CTP wild-type, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.026
-
CTP mutant D211A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.031
-
CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.04
-
CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.048
-
CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.052
-
CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.066
-
N-acetylneuraminate mutant D211A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.068
-
N-acetylneuraminate wild-type, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.11
-
N-acetylneuraminate mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.15
-
CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.27
-
CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.29
-
N-acetylneuraminate mutant R173A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.38
-
N-acetylneuraminate mutant F192A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.78
-
N-acetylneuraminate mutant F193A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 1.6
-
N-acetylneuraminate mutant N175A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 2.6
-
N-acetylneuraminate mutant Q104A, pH 7.5, 22°C Neisseria meningitidis

Metals/Ions

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

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.43 Neisseria meningitidis P0A0Z8
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.7.43 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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.43 CTP + N-acetylneuraminate
-
Neisseria meningitidis diphosphate + CMP-N-acetylneuraminate
-
?
2.7.7.43 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

EC Number Synonyms Comment Organism
2.7.7.43 CMP-Neu5Ac synthetase
-
Neisseria meningitidis
2.7.7.43 CNS
-
Neisseria meningitidis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.43 0.001
-
CTP mutant D211A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.05
-
CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 2.2
-
CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 5
-
CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 7
-
CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 10.2
-
CTP wild-type, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 20
-
CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 25
-
CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.7.43 0.0005
-
N-acetylneuraminate mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.0009
-
N-acetylneuraminate mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.0013
-
CTP mutant Y179A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.0063
-
N-acetylneuraminate mutant F193A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.052
-
N-acetylneuraminate mutant F192A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.072
-
CTP mutant Q104A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.09
-
CTP mutant N175A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.097
-
CTP mutant F193A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.132
-
CTP mutant F192A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.143
-
CTP mutant R173A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.148
-
N-acetylneuraminate wild-type, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.155
-
N-acetylneuraminate mutant N175A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.23
-
N-acetylneuraminate mutant R173A, pH 7.5, 22°C Neisseria meningitidis
2.7.7.43 0.6
-
CTP wild-type, pH 7.5, 22°C Neisseria meningitidis