Summary
The Saccharomyces cerevisiae KEX2 gene encodes the membrane-bound endoprotease yscF, which is responsible for the site-specific endoproteolytic cleavages at pairs of basic amino acid residues in the α-factor precursor. In order to obtain soluble yscF activity, a mutant KEX2 gene lacking 600 bp coding for the C-terminal 200 amino acids was constructed. Expression of the truncated KEX2 gene in yeast led to the secretion of an active soluble yscF protein (yscFs). The soluble yscF protein is able to efficiently cleave heterologous protein precursors in-vitro, as demonstrated for α-factor leader-hIGF1 and α-factor leader-hirudin fusion proteins.
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Seeboth, P.G., Heim, J. In-vitro processing of yeast α-factor leader fusion proteins using a soluble yscF (Kex2) variant. Appl Microbiol Biotechnol 35, 771–776 (1991). https://doi.org/10.1007/BF00169893
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DOI: https://doi.org/10.1007/BF00169893