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Literature summary extracted from

  • Nielsen, M.M.; Suits, M.D.; Yang, M.; Barry, C.S.; Martinez-Fleites, C.; Tailford, L.E.; Flint, J.E.; Dumon, C.; Davis, B.G.; Gilbert, H.J.; Davies, G.J.
    Substrate and metal ion promiscuity in mannosylglycerate synthase (2011), J. Biol. Chem., 286, 15155-15164.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.269 expression in Escherichia coli Rhodothermus marinus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.269 in complex with GDP-Man/Mg2+, GDP/Mg2+, GDP/Mn2+/malonate and lactate, to 2.42-2.9 A resolution. The guanine base of GDP is housed in a hydrophobic pocket between the side chains of Lys9 and Tyr37 on one face and the aliphatic portion of the side chain of Lys76 on the opposing face, with Gln66 accepting hydrogen bonds from the endocyclic N1 and exocyclic N2 Rhodothermus marinus

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.269 D102A completely inactive in the presence of all metals tested Rhodothermus marinus
2.4.1.269 D135A very high Km value for D-glycerate and a slight increase in kcat Rhodothermus marinus
2.4.1.269 D192A inactive Rhodothermus marinus
2.4.1.269 E166A 15-fold increase in Km for glycerate Rhodothermus marinus
2.4.1.269 H217A significant change in metal preference. Mutant is essentially inactive in the presence of Ca2+, whereas the kcat value is reduced 23-fold in the presence of Mg2+ Rhodothermus marinus
2.4.1.269 K76A 3-4-fold increase in kcat coupled with an elevation in Km for D-glycerate and GDP-Man of 15- and 3-fold, respectively Rhodothermus marinus
2.4.1.269 K9A significant increase in kcat, little effect on the Km value for GDP-Man Rhodothermus marinus
2.4.1.269 M229A minor influence on catalytic performance Rhodothermus marinus
2.4.1.269 Q66A substantial increases in kcat and Km for both the acceptor and donor substrates Rhodothermus marinus
2.4.1.269 R218A significant increase in kcat and Km values for GDP-Man, only a modest influence on the Km value for the acceptor substrate Rhodothermus marinus
2.4.1.269 R73A minor influence on catalytic performance Rhodothermus marinus
2.4.1.269 R73G minor influence on catalytic performance Rhodothermus marinus
2.4.1.269 T139A Km value for lactate above 100 mM, modest decrease in kcat but a 1500-fold increase in the Km values for D-glycerate Rhodothermus marinus
2.4.1.269 W189A 3- and 7-fold increase in kcat and Km for D-glycerate, compared with the wild-type enzyme, no influence on utilization of GDP-Man Rhodothermus marinus
2.4.1.269 Y220A 500-fold increase in Km for D-glycerate Rhodothermus marinus
2.4.1.269 Y220F 1500-fold increase in Km for D-glycerate Rhodothermus marinus
2.4.1.269 Y37A significant increase in kcat, little effect on the Km value for GDP-Man Rhodothermus marinus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.269 0.00031
-
GDP-mannose mutant T139A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.001
-
GDP-mannose mutant Q66A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.001
-
GDP-mannose mutant R73A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0014
-
D-glycerate mutant K9A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0017
-
GDP-mannose mutant K9A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0022
-
D-glycerate wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0022
-
D-lactate wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0027
-
GDP-mannose mutant Y37A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0028
-
GDP-mannose mutant M229A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0028
-
GDP-mannose mutant W189A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0029
-
GDP-mannose mutant Y220A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0031
-
GDP-mannose wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0038
-
D-glycerate mutant R73A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0041
-
GDP-mannose mutant R73G, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0041
-
D-glycerate mutant R73G, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0058
-
D-glycerate mutant R218A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0063
-
GDP-mannose mutant D135A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0065
-
GDP-mannose mutant Y220F, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0078
-
D-glycerate mutant M229A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.0096
-
GDP-mannose mutant E166A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.011
-
GDP-mannose mutant K76A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.012
-
GDP-mannose mutant H217A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.015
-
D-glycerate mutant W189A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.017
-
D-glycerate mutant Y37A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.03
-
D-glycerate mutant K76A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.032
-
D-glycerate mutant E166A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.033
-
GDP-mannose mutant R218A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.07
-
D-glycerate mutant Q66A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 1.1
-
D-glycerate mutant Y220A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 1.3
-
D-glycerate mutant D135A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 1.9
-
D-glycerate mutant H217A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 3.3
-
D-glycerate mutant T139A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 3.8
-
D-glycerate mutant Y220F, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.1.269 Ca2+ may partially substitute for Mg2+ Rhodothermus marinus
2.4.1.269 Mg2+ required Rhodothermus marinus
2.4.1.269 Mn2+ may partially substitute for Mg2+ Rhodothermus marinus

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.269 Rhodothermus marinus Q9RFR0 GT78 glycosyltransferase
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.269 GDP-mannose + D-glycerate
-
Rhodothermus marinus 2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP
-
?
2.4.1.269 GDP-mannose + D-lactate
-
Rhodothermus marinus ?
-
?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.269 MGS
-
Rhodothermus marinus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.269 0.017
-
D-glycerate mutant T139A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.017
-
D-lactate wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.018
-
GDP-mannose wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.023
-
D-glycerate mutant E166A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.023
-
D-glycerate mutant Q66A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.028
-
GDP-mannose mutant E166A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.032
-
D-glycerate wild-type, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.037
-
GDP-mannose mutant Y220F, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.042
-
GDP-mannose mutant Y220A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.045
-
GDP-mannose mutant D135A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.047
-
GDP-mannose mutant R73A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.055
-
D-glycerate mutant R73A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.06
-
GDP-mannose mutant R73G, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.06
-
D-glycerate mutant R73G, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.065
-
D-glycerate mutant D135A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.067
-
GDP-mannose mutant W189A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.067
-
D-glycerate mutant Y220A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.068
-
GDP-mannose mutant K76A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.07
-
D-glycerate mutant Y220F, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.075
-
D-glycerate mutant K76A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.077
-
D-glycerate mutant W189A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.082
-
GDP-mannose mutant K9A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.083
-
D-glycerate mutant H217A, pH 7.0, 23°C, presence of 3 mM Mg2+ Rhodothermus marinus
2.4.1.269 0.085
-
GDP-mannose mutant T139A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.115
-
D-glycerate mutant K9A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.127
-
GDP-mannose mutant M229A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.135
-
D-glycerate mutant M229A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.163
-
GDP-mannose mutant Y37A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.183
-
GDP-mannose mutant R218A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.183
-
D-glycerate mutant Y37A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.25
-
D-glycerate mutant R218A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.267
-
D-glycerate mutant H217A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.483
-
GDP-mannose mutant H217A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 0.6
-
D-glycerate wild-type, pH 7.0, 23°C, presence of 3 mM Ca2+ Rhodothermus marinus
2.4.1.269 0.967
-
GDP-mannose mutant Q66A, pH 7.0, 23°C, presence of Mn2+ Rhodothermus marinus
2.4.1.269 1.92
-
D-glycerate wild-type, pH 7.0, 23°C, presence of 3 mM Mg2+ Rhodothermus marinus