cDNA cloning and expression of rat and human protein geranylgeranyltransferase type-I.
Protein geranylgeranyltransferase type-I (GGTase-I) transfers a geranylgeranyl group to the cysteine residue of candidate proteins containing a carboxyl-terminal CAAX (C, cysteine; A, aliphatic amino acid; X, any amino acid) motif in which the "X" residue is leucine. The enzyme is composed of a 48-kilodalton alpha subunit and a 43-kilodalton beta subunit. Peptides isolated from the alpha subunit of GGTase-I were shown to be identical with the alpha subunit of a related enzyme, protein farnesyltransferase. Overlapping cDNA clones containing the complete coding sequence for the beta subunit of GGTase-I were obtained from rat and human cDNA libraries. The cDNA clones from both species each predicted a protein of 377 amino acids with molecular masses of 42.4 kilodaltons (human) and 42.5 kilodaltons (rat). Amino acid sequence comparison suggests that the protein encoded by the Saccharomyces cerevisiae gene CDC43 is the yeast counterpart of the mammalian GGTase-I beta subunit. Co-expression of the GGTase-I beta subunit cDNA together with the alpha subunit of protein farnesyltransferase in Escherichia coli produced recombinant GGTase-I with electrophoretic and enzymatic properties indistinguishable from native GGTase-I.
Zhang
F L
FL
Section of Cell Growth, Regulation and Oncogenesis, Duke University Medical Center, Durham, North Carolina 27710.
Diehl
R E
RE
Kohl
N E
NE
Gibbs
J B
JB
Giros
B
B
Casey
P J
PJ
Omer
C A
CA
D019883:Alkyl and Aryl Transferases; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000235:genetics; D051231:Farnesyltranstransferase; D005656:Fungal Proteins / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005800:Genes, Fungal; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014166:Transferases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D005656:Fungal Proteins; D007527:Isoenzymes; D046911:Macromolecular Substances; D029701:Saccharomyces cerevisiae Proteins; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C083291:geranylgeranyltransferase type-I; D004122:Dimethylallyltranstransferase; D051231:Farnesyltranstransferase; C065203:CDC43 protein, S cerevisiae