公开(公告)号：US07211424B1

公开(公告)日：2007-05-01

申请号：US09830837

申请日：1999-11-04

申请人： Nabil G. Seidah ,  Michel Chretien ,  Mieczyslaw Marcinkiewicz

发明人： Nabil G. Seidah ,  Michel Chretien ,  Mieczyslaw Marcinkiewicz

IPC分类号： C12N9/64 ,  C12N15/57 ,  C12N15/79 ,  C07K1/00 ,  C07K16/00

CPC分类号： C12N9/6424

摘要： Using RT-PCR and degenerate oligonucleotides derived from the active site residues of subtilisin-kexin-like serine proteinases, we have identified a highly conserved and phylogenetically ancestral human, rat and mouse type-I membrane-bound proteinase called subtilisin-kexin-isozyme-1 (SKI-1). Computer data bank searches reveals that human SKI-1 was previously cloned but with no identified function. A SKI-1 processed fragment is secreted in culture media in a soluble form. In vitro studies suggest that SKI-1 is a Ca2+-dependent serine proteinase exhibiting a wide pH optimum for cleavage of proBDNF. Peptides mimicking SKI-1 cleavages sites are also disclosed. SKI-1 prosegment has an ex vivo inhibitory effect on SKI-1 activity. The prosegment is also processed and secreted in culture media. One of its fragments is found tightly associated with the SKI-1 soluble form. Therapeutic applications for SKI-1 inhibitors are disclosed.

摘要翻译： 使用RT-PCR和衍生自枯草杆菌蛋白酶 - 类克隆丝氨酸蛋白酶的活性位点残基的简并寡核苷酸，我们确定了一种高度保守和系统发育的祖先人类，大鼠和小鼠I型膜结合蛋白酶，称为枯草杆菌蛋白酶 - 1（SKI-1）。 计算机数据库搜索显示人类SKI-1以前被克隆，但没有识别功能。 SKI-1处理的片段以可溶形式分泌在培养基中。 体外研究表明，SKI-1是表现出对于proBDNF的切割具有宽pH最适的Ca 2+ 2 +依赖性丝氨酸蛋白酶。 还公开了模拟SKI-1切割位点的肽。 SKI-1 prosegment具有对SKI-1活性的离体抑制作用。 散文也在文化媒体中被处理和分发。 其中一个片段被发现与SKI-1可溶形式紧密相关。 公开了SKI-1抑制剂的治疗应用。