Seeboth P G, Heim J
Ciba-Geigy Ltd., Biotechnology Department, Basel, Switzerland.
Appl Microbiol Biotechnol. 1991 Sep;35(6):771-6. doi: 10.1007/BF00169893.
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.
酿酒酵母KEX2基因编码膜结合的内切蛋白酶yscF，它负责在α-因子前体中碱性氨基酸残基对处进行位点特异性内切蛋白水解切割。为了获得可溶性的yscF活性，构建了一个缺失编码C末端200个氨基酸的600 bp的突变KEX2基因。截短的KEX2基因在酵母中的表达导致分泌出一种活性可溶性yscF蛋白（yscFs）。如α-因子前导肽-hIGF1和α-因子前导肽-水蛭素融合蛋白所示，可溶性yscF蛋白能够在体外有效切割异源蛋白前体。
