Cloned (Comment) | Organism |
---|---|
gene GGPPS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain JM109 | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
H139A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
H139A/R140A | site-directed mutagenesis, inactive monomeric mutant | Saccharomyces cerevisiae |
M111A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
M111E | site-directed mutagenesis, the dimeric/monomeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
M111F | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
N101A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
N101A/N104A | site-directed mutagenesis, the dimeric mutant shows reduced kcat compared to the wild-type | Saccharomyces cerevisiae |
N101A/N104A/Y105A | site-directed mutagenesis, inactive dimeric mutant | Saccharomyces cerevisiae |
N101A/Y105A | site-directed mutagenesis, the dimeric mutant shows reduced kcat compared to the wild-type | Saccharomyces cerevisiae |
N104A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
N104A/Y105A | site-directed mutagenesis, the dimeric mutant shows slightly reduced activity compared to the wild-type | Saccharomyces cerevisiae |
R140A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y105A | site-directed mutagenesis, the dimeric mutant shows altered kinetics compared to the wild-type enzyme | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00061 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.00061 | - |
isopentenyl diphosphate | mutant N101A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0017 | - |
isopentenyl diphosphate | wild-type enzyme, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0025 | - |
(2E,6E)-farnesyl diphosphate | mutant H139A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0025 | - |
isopentenyl diphosphate | mutant M111A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0026 | - |
isopentenyl diphosphate | mutant Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0037 | - |
(2E,6E)-farnesyl diphosphate | wild-type enzyme, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0037 | - |
(2E,6E)-farnesyl diphosphate | mutant M111A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0045 | - |
isopentenyl diphosphate | mutant R140A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0051 | - |
isopentenyl diphosphate | mutant H139A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0055 | - |
(2E,6E)-farnesyl diphosphate | mutant N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0055 | - |
isopentenyl diphosphate | mutant N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0063 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0066 | - |
isopentenyl diphosphate | mutant M111E, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0077 | - |
(2E,6E)-farnesyl diphosphate | mutant Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0081 | - |
isopentenyl diphosphate | mutant N104A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0089 | - |
(2E,6E)-farnesyl diphosphate | mutant R140A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.009 | - |
(2E,6E)-farnesyl diphosphate | mutant M111E, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0113 | - |
isopentenyl diphosphate | mutant M111F, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0146 | - |
(2E,6E)-farnesyl diphosphate | mutant M111F, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0153 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A/N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0292 | - |
isopentenyl diphosphate | mutant N101A/N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0467 | - |
(2E,6E)-farnesyl diphosphate | mutant N104A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.048 | - |
isopentenyl diphosphate | mutant N101A, pH 7.5, 25°C | Saccharomyces cerevisiae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Saccharomyces cerevisiae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
44790 | - |
recombinant mutant H139A/R140A, gel filtration | Saccharomyces cerevisiae |
47820 | - |
recombinant monomeric mutant M111E, gel filtration | Saccharomyces cerevisiae |
72070 | - |
recombinant mutant N101A/Y105A, gel filtration | Saccharomyces cerevisiae |
74740 | - |
recombinant mutant Y105A, gel filtration | Saccharomyces cerevisiae |
75950 | - |
recombinant mutant N101A, gel filtration | Saccharomyces cerevisiae |
76730 | - |
recombinant mutant N104A/Y105A, gel filtration | Saccharomyces cerevisiae |
76990 | - |
recombinant mutant R140A, gel filtration | Saccharomyces cerevisiae |
78690 | - |
recombinant mutant H139A, gel filtration | Saccharomyces cerevisiae |
80350 | - |
recombinant mutant N104A, gel filtration | Saccharomyces cerevisiae |
82300 | - |
recombinant wild-type enzyme, gel filtration | Saccharomyces cerevisiae |
83880 | - |
recombinant mutant N101A/N104A/Y105A, gel filtration | Saccharomyces cerevisiae |
84980 | - |
recombinant mutant N101A/N104A, gel filtration | Saccharomyces cerevisiae |
85920 | - |
recombinant dimeric mutant M111E, gel filtration | Saccharomyces cerevisiae |
88080 | - |
recombinant mutant M111F, gel filtration | Saccharomyces cerevisiae |
92340 | - |
recombinant mutant M111A, gel filtration | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E,6E)-farnesyl diphosphate + isopentenyl diphosphate | Saccharomyces cerevisiae | - |
diphosphate + geranylgeranyl diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | Q12051 | gene GGPPS | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography from Escherichia coli strain JM109, removal of the tag by cleavage with Factor Xa | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(2E,6E)-farnesyl diphosphate + isopentenyl diphosphate | - |
Saccharomyces cerevisiae | diphosphate + geranylgeranyl diphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | dimerization is required for GGPPS activity, resdieus N101 and Y105 form H-bonds with H139 and R140 in the other subunit, and the H-bonds, mainly through N101 for maintaining substrate binding stability and the hydrophobic interaction of M111 in dimer interface, are essential for activity of yeast GGPPS | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
GGPPS | - |
Saccharomyces cerevisiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00013 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.00013 | - |
isopentenyl diphosphate | mutant N101A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0002 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A/N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.0002 | - |
isopentenyl diphosphate | mutant N101A/N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.017 | - |
(2E,6E)-farnesyl diphosphate | mutant N104A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.017 | - |
isopentenyl diphosphate | mutant N104A/Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.02 | - |
(2E,6E)-farnesyl diphosphate | mutant M111E, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.02 | - |
isopentenyl diphosphate | mutant M111E, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.036 | - |
(2E,6E)-farnesyl diphosphate | mutant M111F, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.036 | - |
isopentenyl diphosphate | mutant M111F, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.037 | - |
(2E,6E)-farnesyl diphosphate | wild-type enzyme, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.037 | - |
isopentenyl diphosphate | wild-type enzyme, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.038 | - |
(2E,6E)-farnesyl diphosphate | mutant R140A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.038 | - |
isopentenyl diphosphate | mutant R140A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.04 | - |
(2E,6E)-farnesyl diphosphate | mutant M111A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.04 | - |
isopentenyl diphosphate | mutant M111A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.042 | - |
(2E,6E)-farnesyl diphosphate | mutant H139A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.042 | - |
isopentenyl diphosphate | mutant H139A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.06 | - |
(2E,6E)-farnesyl diphosphate | mutant Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.06 | - |
isopentenyl diphosphate | mutant Y105A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.067 | - |
(2E,6E)-farnesyl diphosphate | mutant N101A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.067 | - |
isopentenyl diphosphate | mutant N101A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.11 | - |
(2E,6E)-farnesyl diphosphate | mutant N104A, pH 7.5, 25°C | Saccharomyces cerevisiae | |
0.11 | - |
isopentenyl diphosphate | mutant N104A, pH 7.5, 25°C | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Saccharomyces cerevisiae |
General Information | Comment | Organism |
---|---|---|
malfunction | yeast geranylgeranyl diphosphate synthase becomes an inactive monomer when the first N-terminal helix involved in dimerization is deleted | Saccharomyces cerevisiae |