Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Rhodothermus marinus |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
D102A | completely inactive in the presence of all metals tested | Rhodothermus marinus |
D135A | very high Km value for D-glycerate and a slight increase in kcat | Rhodothermus marinus |
D192A | inactive | Rhodothermus marinus |
E166A | 15-fold increase in Km for glycerate | Rhodothermus marinus |
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 |
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 |
K9A | significant increase in kcat, little effect on the Km value for GDP-Man | Rhodothermus marinus |
M229A | minor influence on catalytic performance | Rhodothermus marinus |
Q66A | substantial increases in kcat and Km for both the acceptor and donor substrates | Rhodothermus marinus |
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 |
R73A | minor influence on catalytic performance | Rhodothermus marinus |
R73G | minor influence on catalytic performance | Rhodothermus marinus |
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 |
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 |
Y220A | 500-fold increase in Km for D-glycerate | Rhodothermus marinus |
Y220F | 1500-fold increase in Km for D-glycerate | Rhodothermus marinus |
Y37A | significant increase in kcat, little effect on the Km value for GDP-Man | Rhodothermus marinus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00031 | - |
GDP-mannose | mutant T139A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.001 | - |
GDP-mannose | mutant Q66A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.001 | - |
GDP-mannose | mutant R73A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0014 | - |
D-glycerate | mutant K9A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0017 | - |
GDP-mannose | mutant K9A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0022 | - |
D-glycerate | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0022 | - |
D-lactate | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0027 | - |
GDP-mannose | mutant Y37A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0028 | - |
GDP-mannose | mutant M229A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0028 | - |
GDP-mannose | mutant W189A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0029 | - |
GDP-mannose | mutant Y220A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0031 | - |
GDP-mannose | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0038 | - |
D-glycerate | mutant R73A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0041 | - |
GDP-mannose | mutant R73G, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0041 | - |
D-glycerate | mutant R73G, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0058 | - |
D-glycerate | mutant R218A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0063 | - |
GDP-mannose | mutant D135A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0065 | - |
GDP-mannose | mutant Y220F, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0078 | - |
D-glycerate | mutant M229A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.0096 | - |
GDP-mannose | mutant E166A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.011 | - |
GDP-mannose | mutant K76A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.012 | - |
GDP-mannose | mutant H217A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.015 | - |
D-glycerate | mutant W189A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.017 | - |
D-glycerate | mutant Y37A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.03 | - |
D-glycerate | mutant K76A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.032 | - |
D-glycerate | mutant E166A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.033 | - |
GDP-mannose | mutant R218A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.07 | - |
D-glycerate | mutant Q66A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
1.1 | - |
D-glycerate | mutant Y220A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
1.3 | - |
D-glycerate | mutant D135A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
1.9 | - |
D-glycerate | mutant H217A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
3.3 | - |
D-glycerate | mutant T139A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
3.8 | - |
D-glycerate | mutant Y220F, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | may partially substitute for Mg2+ | Rhodothermus marinus | |
Mg2+ | required | Rhodothermus marinus | |
Mn2+ | may partially substitute for Mg2+ | Rhodothermus marinus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodothermus marinus | Q9RFR0 | GT78 glycosyltransferase | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GDP-mannose + D-glycerate | - |
Rhodothermus marinus | 2-O-(alpha-D-mannopyranosyl)-D-glycerate + GDP | - |
? | |
GDP-mannose + D-lactate | - |
Rhodothermus marinus | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MGS | - |
Rhodothermus marinus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.017 | - |
D-glycerate | mutant T139A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.017 | - |
D-lactate | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.018 | - |
GDP-mannose | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.023 | - |
D-glycerate | mutant E166A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.023 | - |
D-glycerate | mutant Q66A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.028 | - |
GDP-mannose | mutant E166A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.032 | - |
D-glycerate | wild-type, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.037 | - |
GDP-mannose | mutant Y220F, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.042 | - |
GDP-mannose | mutant Y220A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.045 | - |
GDP-mannose | mutant D135A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.047 | - |
GDP-mannose | mutant R73A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.055 | - |
D-glycerate | mutant R73A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.06 | - |
GDP-mannose | mutant R73G, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.06 | - |
D-glycerate | mutant R73G, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.065 | - |
D-glycerate | mutant D135A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.067 | - |
GDP-mannose | mutant W189A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.067 | - |
D-glycerate | mutant Y220A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.068 | - |
GDP-mannose | mutant K76A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.07 | - |
D-glycerate | mutant Y220F, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.075 | - |
D-glycerate | mutant K76A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.077 | - |
D-glycerate | mutant W189A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.082 | - |
GDP-mannose | mutant K9A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.083 | - |
D-glycerate | mutant H217A, pH 7.0, 23°C, presence of 3 mM Mg2+ | Rhodothermus marinus | |
0.085 | - |
GDP-mannose | mutant T139A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.115 | - |
D-glycerate | mutant K9A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.127 | - |
GDP-mannose | mutant M229A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.135 | - |
D-glycerate | mutant M229A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.163 | - |
GDP-mannose | mutant Y37A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.183 | - |
GDP-mannose | mutant R218A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.183 | - |
D-glycerate | mutant Y37A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.25 | - |
D-glycerate | mutant R218A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.267 | - |
D-glycerate | mutant H217A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.483 | - |
GDP-mannose | mutant H217A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
0.6 | - |
D-glycerate | wild-type, pH 7.0, 23°C, presence of 3 mM Ca2+ | Rhodothermus marinus | |
0.967 | - |
GDP-mannose | mutant Q66A, pH 7.0, 23°C, presence of Mn2+ | Rhodothermus marinus | |
1.92 | - |
D-glycerate | wild-type, pH 7.0, 23°C, presence of 3 mM Mg2+ | Rhodothermus marinus |