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Appl Microbiol Biotechnol

Title:		In-vitro processing of yeast alpha-factor leader fusion proteins using a soluble yscF (Kex2) variant
Author(s):		Seeboth PG; Heim J;
Address:		"Ciba-Geigy Ltd., Biotechnology Department, Basel, Switzerland"
Journal Title:		Appl Microbiol Biotechnol
Year:		1991
Volume:		35
Issue:		6
Page Number:		771 - 776
DOI:		10.1007/BF00169893
ISSN/ISBN:		0175-7598 (Print) 0175-7598 (Linking)
Abstract:		"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 alpha-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 alpha-factor leader-hIGF1 and alpha-factor leader-hirudin fusion proteins"
Keywords:		Colorimetry Fermentation Fungal Proteins/*metabolism Hirudins/metabolism Immunoblotting Insulin-Like Growth Factor I/metabolism Mating Factor Mutation Peptides/metabolism Plasmids *Proprotein Convertases Protein Precursors/metabolism Recombinant Fusion Pr;
Notes:		"MedlineSeeboth, P G Heim, J eng Germany 1991/09/01 Appl Microbiol Biotechnol. 1991 Sep; 35(6):771-6. doi: 10.1007/BF00169893"