Characterization of an endopeptidase involved in pre-protein processing

A. W. Strauss, M. Zimmerman, I. Boime, B. Ashe, R. A. Mumford, A. W. Alberts

Research output: Contribution to journalArticle

46 Scopus citations

Abstract

Proteolytic removal of the pre-segment from growing nascent chains of pre-human placental lactogen (hPL) occurred during in vitro translation mRNA if crude membranes derived from ascites lysates dog pancreas, or rat liver rough endoplasmic reticulum were added to the translation mixtures. The cotranslational proteolytic event was inhibited by the peptide protease inhibitor, chymostatin, but not by leupeptin, antipain, or elastatinal. The proteases involved in cleavage were solubilized with detergent and converted completed pre-hPL to hPL (post-translational processing). Direct assay of the solubilized membranes, with synthetic fluorogenic aminocoumarin peptide substrates, revealed no significant tryptic or elastase-like activity, but activity against a chymotrypsin substrate [(succinyl-Ala-Ala-Phe)-7-amino-4-methylcoumarin] was found. This activity was dependent upon both an endopeptidase and an aminopeptidase. Although bestatin inhibited the aminopeptidase activity, it had no effect on the endopeptidase or on post-translational cleavage. Although this endopeptidase cleaved on the COOH side of an alanine residue, it was not inhibited by elastatinal. However, it was inhibited by levels of chymostatic and by some serine protease inhibitors.

Original languageEnglish
Pages (from-to)4225-4229
Number of pages5
JournalUnknown Journal
Volume76
Issue number9
DOIs
StatePublished - Jan 1 1979

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    Strauss, A. W., Zimmerman, M., Boime, I., Ashe, B., Mumford, R. A., & Alberts, A. W. (1979). Characterization of an endopeptidase involved in pre-protein processing. Unknown Journal, 76(9), 4225-4229. https://doi.org/10.1073/pnas.76.9.4225