Carboxy-terminal CFFL-sequence-specific monomeric protein geranylgeranyltransferase I from the eyes of the shrimp Penaeus japonicus.
Protein geranylgeranyltransferase I from the eyes of Penaeus japonicus geranylgeranylates predominantly the sequence CFFL and Drosophila-specific Ras1 carboxyl termini, with the sequence CKML, as well as mammalian-specific G gamma carboxyl termini, with the sequence CAIL, but not the protein farnesyltransferase-specific sequence CVLS. The purified protein geranylgeranyltransferase I from shrimp was evidenced by immunoblotting and polyacrylamide gel electrophoresis under denaturing conditions to consist of single subunit of Mr 66,000 +/- 500. Since the active protein geranylgeranyltransferase I was found to have a relative mass of 67,000 +/- 1,000, the purified enzyme was deduced to be a monomer. The enzyme had an optimal pH of 8.0 with 100 mM Tris as the buffer and a K(m) of 7 +/- 2 microM with the synthetic peptide KCFFL as the substrate. The enzyme was inhibited by Zn++ and Mg++ ions at micromolar concentrations.
Lin
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Division of Biochemistry and Molecular Sciences, Academia Sinica, Taipei, Taiwan. zonnc@sinica.edu.tw
Chuang
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D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005123:Eye / Q000201:enzymology; D007700:Kinetics; D008970:Molecular Weight; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D033561:Penaeidae / Q000201:enzymology*; D017368:Protein Prenylation; D013379:Substrate Specificity
D009842:Oligopeptides; D019883:Alkyl and Aryl Transferases; C083291:geranylgeranyltransferase type-I