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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Mammalian subtilisin/kexin isozyme SKI-1: A widely expressed proprotein convertase with a unique cleavage specificity and cellular localization
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Mammalian subtilisin/kexin isozyme SKI-1: A widely expressed proprotein convertase with a unique cleavage specificity and cellular localization

机译：哺乳动物枯草杆菌蛋白酶/ kexin同工酶SKI-1：广泛表达的前蛋白转化酶，具有独特的裂解特异性和细胞定位

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摘要

Using reverse transcriptase-PCR and degen- erate oligonucleotides derived from the active-site residues of subtilisin/kexin-like serine proteinases, we have identified a highly conserved and phylogenetically ancestral human, raL and mouse type I membrane-bound proteinase called subtili- sin/kexin-isozyme-1 (SKI-I). Computer databank searches re- veal that human SKI-1 was cloned previously but with no identified function. In situ hybridization demonstrates that SKI-l mRNA is present in most tissues and cells. Cleavage specificity studies show that SKI-I generates a 28-kDa prod- uct from the 32-kDa brain-derived neurotrophic factor pre- cursor cleaving at an RGLT 1 SL bond. In the endoplasmic reticulum of either LoVo or HK293 cells, ProSKI-1 is pro- cessed into two membrane-bound forms of SKI-1 (120 and 106 cha) differing by the nature of their N-glycosylation. Late along the secretory pathway some of the membrane-hound enzyme is shed into the medium as a 98-kDa form. lmmuno- cytochemical analysis of stably transfected HK293 cclls shows Ihat SKI-1 is present in the Golgi apparatus and within small punctate structures reminiscent of endosomes. In vitro studies suggest that SKI-1 is a Ca2+-dependent serine proteinase exhibiting a wide PH optimum for cleavage of pro-brain- derived neurotrophic factor.
机译：使用逆转录酶-PCR和衍生自枯草杆菌蛋白酶/ kexin样丝氨酸蛋白酶活性位点的简并寡核苷酸，我们鉴定出了一种高度保守的，系统发育的人，raL和小鼠I型膜结合蛋白酶，称为subtili- sin / kexin-isozyme-1（SKI-1）。计算机数据库搜索显示，以前克隆了人SKI-1，但没有确定的功能。原位杂交表明SKI-1 mRNA存在于大多数组织和细胞中。切割特异性研究表明，SKI-1从32kDa的脑源性神经营养因子前体在RGLT 1 SL键处裂解产生了28kDa的产物。在LoVo或HK293细胞的内质网中，ProSKI-1被加工成两种膜结合形式的SKI-1（120和106 cha），它们的N-糖基化性质不同。沿着分泌途径的后期，一些膜猎犬以98-kDa的形式进入培养基。稳定转染的HK293细胞的免疫细胞化学分析显示，Ihat SKI-1存在于高尔基体中以及让人联想到内体的小点状结构中。体外研究表明，SKI-1是一种Ca2 +依赖性丝氨酸蛋白酶，对裂解前脑源性神经营养因子具有最佳的PH值。

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