Hatsuzawa K, Hosaka M, Nakagawa T, Nagase M, Shoda A, Murakami K, Nakayama K
Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
J Biol Chem. 1990 Dec 25;265(36):22075-8.
We have cloned and sequenced a mouse cDNA encoding the 793-residue amino acid sequence of furin, which is a protein homologous to the yeast Kex2 protease. The entire sequence is 94% identical to that of human furin, and it contains the 289-residue sequence of the subtilisin-like catalytic domain. Within this region, 99, 64, and 53% of the amino acids are identical to those of human furin, human PC2 (the other mammalian Kex2-like protein), and yeast Kex2, respectively. It has been proposed that furin is a mammalian prohormone processing enzyme which cleaves precursors at paired basic amino acids, based on the fact that the Kex2 protease is responsible for processing of alpha-mating factor and killer toxin precursors at dibasic sites. However, Northern blot analysis has revealed that a furin mRNA transcript is present in all tested mouse tissues and culture cell lines, including those known not to process prohormones. Moreover, when furin and a prohormone, prorenin, have been coexpressed in mammalian cells by DNA transfection, no processing has been observed. These observations fail to show a role for furin, a Kex2-like mammalian protease, in prohormone processing.
我们已经克隆并测序了一个小鼠cDNA,它编码弗林蛋白酶的793个氨基酸残基序列,弗林蛋白酶是一种与酵母Kex2蛋白酶同源的蛋白质。其整个序列与人类弗林蛋白酶的序列有94%的同一性,并且它包含枯草杆菌蛋白酶样催化结构域的289个氨基酸残基序列。在这个区域内,分别有99%、64%和53%的氨基酸与人类弗林蛋白酶、人类PC2(另一种哺乳动物Kex2样蛋白)和酵母Kex2的氨基酸相同。基于Kex2蛋白酶负责在双碱性位点加工α-交配因子和杀伤毒素前体这一事实,有人提出弗林蛋白酶是一种哺乳动物激素原加工酶,它在成对的碱性氨基酸处切割前体。然而,Northern印迹分析显示,在所有测试的小鼠组织和培养细胞系中都存在弗林蛋白酶mRNA转录本,包括那些已知不加工激素原的组织和细胞系。此外,当通过DNA转染在哺乳动物细胞中共表达弗林蛋白酶和一种激素原——肾素原时,未观察到加工现象。这些观察结果未能表明Kex2样哺乳动物蛋白酶弗林蛋白酶在激素原加工中起作用。