Activating Compound | Comment | Organism | Structure |
---|---|---|---|
Triton X-100 | activating at 0.1% | Saccharomyces cerevisiae |
Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escheichia coli strain BL21(DE3) | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
native and selenomethionine-labeled enzymes, hanging drop vapour diffusion method, mixing of 0.002 ml of the GGPP solution with 1012 mg/ml protein in 25 mM Tris-HCl, pH 7.5, 150 mM NaCl, and 0.1% Triton X-100 with 0.002 ml of the mother liquor containing 0.08 M CH3COONa, 0.145 M (NH4)2SO4, 13% polyethylene glycol 4000, 7-9% glycerol, and 7-9% 1,2-propanediol, and equilibrating with 0.5 ml of the mother liquor, 7 days room temperature, X-ray diffraction structure determination and analysis at 1.98 A resolution | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
F108A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
F108A/H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
L135A/H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
additional information | deletion of the first 9 or 17 amino acids caused the dissociation of dimer into monomer, the DELTA1-17 mutant shows abolished enzyme activity | Saccharomyces cerevisiae |
S71Y | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y107A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y107A/F108A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y107A/F108A//L135A/H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y107A/F108A/H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
Y107A/H139A | site-directed mutagenesis, mutant activity, substrate specificity, and product distribution compared to the wild-type enzyme | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes | Saccharomyces cerevisiae | |
0.0004 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/H139A | Saccharomyces cerevisiae | |
0.0008 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
0.001 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/L135A/H139A | Saccharomyces cerevisiae | |
0.0011 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/H139A | Saccharomyces cerevisiae | |
0.0017 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A | Saccharomyces cerevisiae | |
0.0021 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A | Saccharomyces cerevisiae | |
0.0025 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant H139A | Saccharomyces cerevisiae | |
0.0026 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A | Saccharomyces cerevisiae | |
0.0027 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A | Saccharomyces cerevisiae | |
0.0027 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant L135A/H139A | Saccharomyces cerevisiae | |
0.0032 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
0.0033 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A | Saccharomyces cerevisiae | |
0.0047 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A | Saccharomyces cerevisiae | |
0.0051 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A/H139A | Saccharomyces cerevisiae | |
0.0051 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant H139A | Saccharomyces cerevisiae | |
0.017 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant L135A/H139A | Saccharomyces cerevisiae | |
0.051 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/L135A/H139A | Saccharomyces cerevisiae | |
0.097 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/H139A | Saccharomyces cerevisiae | |
0.136 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A/H139A | Saccharomyces cerevisiae | |
0.267 | - |
isopentenyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/H139A | Saccharomyces cerevisiae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans,trans-farnesyl diphosphate + isopentenyl diphosphate | Saccharomyces cerevisiae | - |
diphosphate + geranylgeranyl diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(2E,6E)-farnesyl diphosphate + isopentenyl diphosphate = diphosphate + geranylgeranyl diphosphate | reaction mechanism and residues involved in substrate specificity and product chain lengt determination, overview | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans,trans-farnesyl diphosphate + isopentenyl diphosphate | - |
Saccharomyces cerevisiae | diphosphate + geranylgeranyl diphosphate | - |
? | |
trans,trans-farnesyl diphosphate + isopentenyl diphosphate | - |
Saccharomyces cerevisiae | diphosphate + geranylgeranyl diphosphate | product distribution of the wild-type and mutant enzymes, overview, mechanism of product chain length determination | ? |
Subunits | Comment | Organism |
---|---|---|
More | the enzyme structure is composed entirely of 15 alpha-helices joined by connecting loops and is arranged with alpha-helices around a large central cavity, the N-terminal 17 amino acids, 9-amino acid helix A and the following loop, of this GGPPs protrude from the helix core into the other subunit and contribute to the tight dimer formation, in each subunit, an elongated hydrophobic crevice surrounded by D, F, G, H, and I alpha-helices contains twoDDXXDmotifs at the top for substrate binding with one Mg2+ coordinated by Asp75, Asp79, and four water molecules. It is sealed at the bottom with three large residues of Tyr107, Phe108, and His139 | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
short-chain type-III GGPP | - |
Saccharomyces cerevisiae |
type-III geranylgeranyl pyrophosphate synthase | - |
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 |
---|---|---|---|---|---|
1.6 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A/H139A | Saccharomyces cerevisiae | |
2.5 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant wild-type enzyme | Saccharomyces cerevisiae | |
2.5 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/H139A | Saccharomyces cerevisiae | |
3.9 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/H139A | Saccharomyces cerevisiae | |
4.2 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant H139A | Saccharomyces cerevisiae | |
4.7 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant F108A | Saccharomyces cerevisiae | |
4.9 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A | Saccharomyces cerevisiae | |
5.6 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A | Saccharomyces cerevisiae | |
6.4 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant L135A/H139A | Saccharomyces cerevisiae | |
8.4 | - |
trans,trans-farnesyl diphosphate | pH 7.5, 25°C, recombinant mutant Y107A/F108A/L135A/H139A | Saccharomyces cerevisiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Saccharomyces cerevisiae |