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首页> 外文期刊>Protein Expression and Purification >LEKTI domain 15 is a functional Kazal-type proteinase inhibitor
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LEKTI domain 15 is a functional Kazal-type proteinase inhibitor

机译：LEKTI域15是功能性的Kazal型蛋白酶抑制剂

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摘要

The multidomain proteinase inhibitor LEKTI (lympho-epithelial Kazal-type related inhibitor) consists of 15 potential serine proteinase inhibitory domains. In various diseases such as the severe skin disorder Netherton syndrome as well as atopy, defects in the gene encoding LEKTI have been identified that generate premature termination codons of translation, suggesting a specific role of the COOH-terminal part of LEKTI in healthy individuals. We overexpressed and purified a sequence comprising the 15th domain of LEKTI for further characterisation. Here, we present a high yield expression system for recombinant production and efficient purification of LEKTI domain 15 as a highly soluble protein with a uniform disulfide pattern that is identical to that of other known Kazal-type inhibitors. Also, the expected P1P1' site was confirmed. LEKTI domain 15 is a well-structured protein as verified by circular dichroisin (CD) spectroscopy and a tight-binding and stable inhibitor of the serine proteinase trypsin. These findings confirm the designation of domain 15 as a proteinase inhibitor of the Kazal family. (c) 2007 Elsevier Inc. All rights reserved.
机译：多域蛋白酶抑制剂LEKTI（淋巴上皮Kazal型相关抑制剂）由15个潜在的丝氨酸蛋白酶抑制域组成。在各种疾病中，例如严重的皮肤病Netherton综合征以及特应性疾病，已经识别出编码LEKTI的基因中的缺陷，该缺陷会产生翻译的过早终止密码子，提示LEKTI的COOH末端部分在健康个体中起特定作用。我们过表达并纯化了包含LEKTI第15个结构域的序列，以进行进一步表征。在这里，我们提出了一种高产量的表达系统，用于重组生产和高效纯化LEKTI域15，这是一种具有均匀二硫键模式的高可溶性蛋白，与其他已知的Kazal型抑制剂相同。此外，确认了预期的P1P1'网站。 LEKTI域15是一种结构良好的蛋白质，已通过圆二色球蛋白（CD）光谱验证，是丝氨酸蛋白酶胰蛋白酶的紧密结合且稳定的抑制剂。这些发现证实了结构域15被指定为Kazal家族的蛋白酶抑制剂。 （c）2007 Elsevier Inc.保留所有权利。

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