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Literature summary for 2.5.1.59 extracted from

  • Kim, H.; Yang, C.H.
    Active site determination of yeast geranylgeranyl protein transferase type I expressed in Escherichia coli (1999), Eur. J. Biochem., 265, 105-111.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
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

Protein Variants

Protein Variants Comment Organism
D140N alpha-subunit, increased Km Saccharomyces cerevisiae
H145D alpha-subunit, increased Km Saccharomyces cerevisiae
H216D beta-subunit, KM of geranylgeranyl diphosphate increased 12fold Saccharomyces cerevisiae
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
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
N282A alpha-subunit, increased Km Saccharomyces cerevisiae
R166I beta-subunit, KM of geranylgeranyl diphosphate increased 29fold, no forming of geranygeranyl diphosphate Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
diethyl dicarbonate 80% loss of activity at 5 mM Saccharomyces cerevisiae
Phenylglyoxal 80% loss of activity, inactivation of inhibition in the presence of geranylgeranyl diphosphate Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information comparison of Km of wild-type and mutant enzyme Saccharomyces cerevisiae

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
geranylgeranyl diphosphate + protein-cysteine Saccharomyces cerevisiae
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S-geranylgeranyl-protein + diphosphate
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?

Organism

Organism UniProt Comment Textmining
Saccharomyces cerevisiae
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-
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Posttranslational Modification

Posttranslational Modification Comment Organism
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

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes Saccharomyces cerevisiae

Reaction

Reaction Comment Organism Reaction ID
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
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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
geranylgeranyl diphosphate + protein-cysteine
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Saccharomyces cerevisiae S-geranylgeranyl-protein + diphosphate
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?
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
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?
geranylgeranyl diphosphate + protein-cysteine prenylation, substrates are Rho, Rac, most trimeric G protein gamma subunits Saccharomyces cerevisiae S-geranylgeranyl-protein + diphosphate
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?
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
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?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
additional information comparison of kcat of wild-type and mutant enzyme Saccharomyces cerevisiae