Cloned (Comment) | Organism |
---|---|
wild-type and mutant geranyl diphosphate methyltransferase expression as His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Streptomyces coelicolor |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His6-tagged wild-type enzyme in complex with S-adenosyl-L-methionine and monoterpenoid ligands geranyl-S-thiolodiphosphate, a substrate analogue, or geranyl diphosphate, sitting drop vapor diffusion method, 4°C, mixing of 0.001 ml protein solution conatining 6 mg/mL enzyme, 50 mM PIPES, pH 6.7, 20% glycerol, 5 mM BME, 15 mM MgCl2, 100 mM NaCl, 10 mM S-adenosyl-L-methionine, and 1 mM monoterpenoid ligand with 0.001 ml of precipitant solution containing 100 mM HEPES, pH 7.5, 25% polyethylene glycol 3350, and 200 mM (NH4)2SO4 for cocrystallization with geranyl-S-thiolodiphosphate, 100 mM Bis-Tris, pH 6.5, 25% polyethylene glycol 3350, and 200 mM (NH4)2SO4 for cocrystallization with geranyl diphosphate and equilibrated against a 0.1 ml reservoir of precipitant solution at 21°C, 1 day, X-ray diffraction structure determination and analysis at 2.05 A resolution | Streptomyces coelicolor |
Protein Variants | Comment | Organism |
---|---|---|
Y51F | site-directed mutagenesism the utant shows a 2fold reduction in kcat and no effect on KM for either substrate compared to the wild-type enzyme | Streptomyces coelicolor |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + geranyl diphosphate | Streptomyces coelicolor | - |
S-adenosyl-L-homocysteine + (E)-2-methylgeranyl diphosphate | - |
? | |
S-adenosyl-L-methionine + geranyl diphosphate | Streptomyces coelicolor A3(2) | - |
S-adenosyl-L-homocysteine + (E)-2-methylgeranyl diphosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces coelicolor | Q9F1Y5 | - |
- |
Streptomyces coelicolor A3(2) | Q9F1Y5 | - |
- |
Purification (Comment) | Organism |
---|---|
soluble recombinant His6-tagged wild-type and mutant geranyl diphosphate methyltransferase from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Streptomyces coelicolor |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + geranyl diphosphate = S-adenosyl-L-homocysteine + (E)-2-methylgeranyl diphosphate | catalytic reaction mechanism, detailed overview | Streptomyces coelicolor |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + geranyl diphosphate | - |
Streptomyces coelicolor | S-adenosyl-L-homocysteine + (E)-2-methylgeranyl diphosphate | - |
? | |
S-adenosyl-L-methionine + geranyl diphosphate | - |
Streptomyces coelicolor A3(2) | S-adenosyl-L-homocysteine + (E)-2-methylgeranyl diphosphate | - |
? |
Subunits | Comment | Organism |
---|---|---|
hexamer | hexameric quaternary structure of geranyl diphosphate methyltransferase in the crystal, overview | Streptomyces coelicolor |
Synonyms | Comment | Organism |
---|---|---|
geranyl diphosphate C-methyltransferase | - |
Streptomyces coelicolor |
GPPMT | - |
Streptomyces coelicolor |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Streptomyces coelicolor |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Streptomyces coelicolor |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Streptomyces coelicolor |
General Information | Comment | Organism |
---|---|---|
metabolism | geranyl diphosphate C-methyltransferase is the methyltransferase that modifies an acyclic isoprenoid diphosphate, geranyl diphosphate, to yield a noncanonical acyclic allylic diphosphate product, 2-methylgeranyl diphosphate, which serves as the substrate for a subsequent cyclization reaction catalyzed by a terpenoid cyclase, methylisoborneol synthase | Streptomyces coelicolor |
physiological function | geranyl diphosphate C-methyltransferase is the methyltransferase that modifies an acyclic isoprenoid diphosphate, geranyl diphosphate, to yield a noncanonical acyclic allylic diphosphate product, 2-methylgeranyl diphosphate, which serves as the substrate for a subsequent cyclization reaction catalyzed by a terpenoid cyclase, methylisoborneol synthase | Streptomyces coelicolor |