1. Enhanced peptide secretion by gene disruption ofCYM1, a novel protease inSaccharomyces cerevisiae.
- Author
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Jønson, Lars, Rehfeld, Jens F., and Johnsen, Anders H.
- Subjects
SACCHAROMYCES cerevisiae ,SOMATOTROPIN ,NEUROPEPTIDES ,PROTEINS ,PEPTIDES ,BIOMOLECULES ,SACCHAROMYCES - Abstract
Saccharomyces cerevisiaeis a widely used host in the production of therapeutic peptides and proteins. Here we report the identification of a novel endoprotease inS. cerevisiae.It is encoded by theCYM1gene and is specific for the C-terminus of basic residues of heterologously expressed peptides. Gene disruption ofCYM1not only reduced the intracellular proteolysis, but also enhanced the secretion of heterologously expressed peptides such as growth hormone, pro-B-type natriuretic peptide and pro-cholecystokinin. Cym1p resembles metalloendoproteases of the pitrilysin family with the HXXEH(X)E(71–77)catalytic domain as seen in insulysin, nardilysin and human metalloprotease 1. It is a nuclear encoded protease that localizes to mitochondria without a hydrophobic N-terminal signal sequence or a C-terminal tail-anchor. The protease does not require post-translational processing prior to activation and it contains cytosolic activity that processes peptides designated for the secretory pathway prior to translocation into the endoplasmic reticulum. [ABSTRACT FROM AUTHOR]
- Published
- 2004
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