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1. 发明授权

US06750034B1 DNA encoding human serine protease D-G 有权
标题翻译：编码人丝氨酸蛋白酶D-G的DNA
公开(公告)号：US06750034B1
公开(公告)日：2004-06-15
申请号：US09607745
申请日：2000-06-30
申请人： Andrew L. Darrow , Jenson (Jian-Shen) Qi , Patricia Andrade-Gordon
发明人： Andrew L. Darrow , Jenson (Jian-Shen) Qi , Patricia Andrade-Gordon
IPC分类号： C12Q134
CPC分类号： C12N9/6454 , A61K38/00 , Y10T436/24
摘要： Here we describe the molecular identification of a cDNA encoding a novel serine protease we have termed D-G. The deduced amino acid sequence, and it's alignment with other well characterized serine proteases clearly indicates that it is a member of the S1 serine protease family. We have found that the protease D-G mRNA is widely expressed in several tissues throughout the body including epidermis, fibroblasts, keratinocytes, colon, small intestine, stomach, lung, kidney, bone marrow, lymph node, thymus, ovary, prostate, uterus and spinal cord. Interestingly, this protease contains a hydrophobic stretch of amino acids which is a putative transmembrane near the NH2-terminus. Thus, this serine protease is thought to be synthesized as a type II integral protein. We expressed a soluble form of this novel human protease by inserting the portion of the protease D-G cDNA, encoding the catalytic domain, in a zymogen activation construct designed to permit the generic activation of heterologous serine protease catalytic domains. The result is an active preparation of protease D-G that has an activity against a subset of amidolytic substrates. This enzymatically active protease D-G preparation is now amenable to further biochemical analyses for the identification of physiological substrates as well as specific inhibitors.
摘要翻译：这里我们描述了编码我们称为D-G的新型丝氨酸蛋白酶的cDNA的分子鉴定。推导的氨基酸序列，与其他良好表征的丝氨酸蛋白酶的比对清楚地表明它是S1丝氨酸蛋白酶家族的成员。我们已经发现蛋白酶DG mRNA在全身的几种组织中广泛表达，包括表皮，成纤维细胞，角质形成细胞，结肠，小肠，胃，肺，肾，骨髓，淋巴结，胸腺，卵巢，前列腺，子宫和脊髓绳子有趣的是，该蛋白酶含有氨基酸的疏水延伸，氨基酸是NH2末端附近的推定跨膜。因此，认为该丝氨酸蛋白酶被合成为II型整合蛋白。我们通过将编码催化结构域的蛋白酶D-G cDNA的部分插入设计用于允许异源丝氨酸蛋白酶催化结构域的通用活化的酶原激活构建体中来表达这种新型人类蛋白酶的可溶形式。其结果是蛋白酶D-G的活性制备，其具有对酰氨基底物的子集的活性。这种酶活性蛋白酶D-G制剂现在适合进一步的生物化学分析用于鉴定生理底物以及特异性抑制剂。

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