A microsomal endoprotease that specifically cleaves isoprenylated peptides.
A microsomal enzymatic activity is described that can specifically cleave the tetrapeptide N-acetyl-S-farnesyl-L-Cys-L-Val-L-Ile-L-Ser between the isoprenylated cysteine residue and the valine residue. Km and Vmax values are measured as 5.8 microM and 251 pmol/min per mg of protein, respectively. Proteolytic cleavage of the substrate is stereospecific because the substitution of a farnesylated D-cysteine residue for the L-amino acid leads to the abolition of substrate activity. A free carboxyl-terminal group is also required for substrate activity because methyl esterification renders the substrate inert. The tripeptide N-acetyl-S-farnesyl-L-Cys-L-Val-L-Ile and the dipeptide N-acetyl-S-farnesyl-L-Cys-L-Val are also hydrolyzed by the protease. Again, stereospecificity is observed at the isoprenylated residue. Hydrolysis of the farnesylated tetrapeptide is not inhibited by a 5-fold excess of the nonfarnesylated tetrapeptide, suggesting that isoprenylation is important for substrate activity. This activity is probably the same as the proteolytic activity proposed to cleave isoprenylated proteins terminating in a Cys-Ali-Ali-Xaa motif, where Ali refers to aliphatic amino acid. These proteins include the ras family of G proteins and the heterotrimeric G proteins. Proteolytic maturation of these essential isoprenylated signal-transducing elements is a key step in their activation.
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Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
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D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003545:Cysteine / Q000737:chemistry; D010450:Endopeptidases / Q000378:metabolism*; D005204:Farnesol; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D013379:Substrate Specificity
D009842:Oligopeptides; D005204:Farnesol; D010450:Endopeptidases; D003545:Cysteine