EC Number | Cloned (Comment) | Organism |
---|---|---|
2.5.1.59 | alpha-subunit encoded by RAM2 cloned to the pFC vector, which has an chloroamphenicol resistance gene and beta-subunit encoded by CAL1 cloned to the pFlag vector, which has an ampicillin resistance gene, these two recombinant enzymes are cotransformed and expressed in Escherichia coli | Saccharomyces cerevisiae |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
2.5.1.59 | D140N | alpha-subunit, increased Km | Saccharomyces cerevisiae |
2.5.1.59 | H145D | alpha-subunit, increased Km | Saccharomyces cerevisiae |
2.5.1.59 | H216D | beta-subunit, KM of geranylgeranyl diphosphate increased 12fold | Saccharomyces cerevisiae |
2.5.1.59 | additional information | comparison of substrate specificity, using GST-CAIL and geranylgeranyl diphosphate: best substrate of wild-type enzyme, using GST-CVIM and geranylgeranyl diphosphate: best substrate of mutant beta H216D, using GST-CAIL and farnesyl diphosphate: best substrate of mutant betaR166I | Saccharomyces cerevisiae |
2.5.1.59 | additional information | seven different mutations in CAL1/CDC43 gene: cal1-1 and cdc43-2 to cdc43-7, all of mutants possess reduced activity and none show temperature-sensitive enzymatic activities, but all of them show temperature-sensitive growth phenotypes | Saccharomyces cerevisiae |
2.5.1.59 | N282A | alpha-subunit, increased Km | Saccharomyces cerevisiae |
2.5.1.59 | R166I | beta-subunit, KM of geranylgeranyl diphosphate increased 29fold, no forming of geranygeranyl diphosphate | Saccharomyces cerevisiae |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
2.5.1.59 | diethyl dicarbonate | 80% loss of activity at 5 mM | Saccharomyces cerevisiae | |
2.5.1.59 | Phenylglyoxal | 80% loss of activity, inactivation of inhibition in the presence of geranylgeranyl diphosphate | Saccharomyces cerevisiae |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.5.1.59 | additional information | - |
additional information | comparison of Km of wild-type and mutant enzyme | Saccharomyces cerevisiae |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine | Saccharomyces cerevisiae | - |
S-geranylgeranyl-protein + diphosphate | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
2.5.1.59 | Saccharomyces cerevisiae | - |
- |
- |
EC Number | Posttranslational Modification | Comment | Organism |
---|---|---|---|
2.5.1.59 | side-chain modification | chemical modification with phenylglyoxal of arginine residue: 80% loss of activity in 30 min, chemical modification with diethyl dicarbonate of histidine residue: 80% loss of activity at 5 mM | Saccharomyces cerevisiae |
EC Number | Purification (Comment) | Organism |
---|---|---|
2.5.1.59 | recombinant wild-type and mutant enzymes | Saccharomyces cerevisiae |
EC Number | Reaction | Comment | Organism | Reaction ID |
---|---|---|---|---|
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine = S-geranylgeranyl-protein + diphosphate | catalyses the formation of a thioether linkage between the C-1 atom of the geranylgeranyl group and a cysteine residue fourth from the C-terminus of the protein, these protein acceptors have the C-terminal sequence CA1A2X, where the terminal residue, X, is preferably leucine, but serine, methionine, alanine or glutamine makes the protein a substrate for EC 2.5.1.58, the enzymes are relaxed in specificity for A1, but cannot act if A2 is aromatic, known targets of this enzyme include most g-subunits of heterotrimeric G proteins and Ras-related GTPases such as members of the Ras and Rac/Rho families | Saccharomyces cerevisiae | |
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine = S-geranylgeranyl-protein + diphosphate | this enzyme, along with protein farnesyltransferase, EC 2.5.1.58 and protein geranylgeranyltransferase type II, EC 2.5.1.60, constitutes the protein prenyltransferase family of enzymes | Saccharomyces cerevisiae |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine | - |
Saccharomyces cerevisiae | S-geranylgeranyl-protein + diphosphate | - |
? | |
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine | enzyme requires that protein substrates contain a Cys residue fourth from the C terminus, protein substrate motif: Cys-Leu | Saccharomyces cerevisiae | S-geranylgeranyl-protein + diphosphate | - |
? | |
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine | prenylation, substrates are Rho, Rac, most trimeric G protein gamma subunits | Saccharomyces cerevisiae | S-geranylgeranyl-protein + diphosphate | - |
? | |
2.5.1.59 | geranylgeranyl diphosphate + protein-cysteine | enzyme requires that protein substrates contain a Cys residue fourth from the C terminus, protein substrate motif: Cys-aliphatic-aliphatic-X | Saccharomyces cerevisiae | S-geranylgeranyl-protein + diphosphate | - |
? |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
2.5.1.59 | additional information | - |
additional information | comparison of kcat of wild-type and mutant enzyme | Saccharomyces cerevisiae |