Characterization of an intracellular cystatin homolog from the tick Haemaphysalis longicornis
Characterization of an intracellular cystatin homolog from the tick Haemaphysalis longicornis
复制标题
DOI：
10.1016/j.vetpar.2008.10.086
复制
发表时间：
2009-03-09
影响因子：
2.6
通讯作者：
Fujisaki, Kozo
中科院分区：
文献类型：
--
作者：
Zhou, Jinlin;Liao, Min;Fujisaki, Kozo
胱抑素是木瓜蛋白酶样半胱氨酸蛋白酶的紧密结合抑制剂。根据氨基酸序列分析，半胱氨酸蛋白酶抑制剂可分为三个密切相关的家族1、家族2和家族3。其中，只有家族I半胱氨酸蛋白酶抑制剂是细胞内的。本文报道了长角血蜱（Haemaphysalis longicornis，Hlcyst-1）中编码第1家族半胱氨酸蛋白酶抑制剂的基因。其cDNA全长为437 bp，包括一个完整的开放阅读框，编码98个氨基酸。序列分析表明，它与已知的家族1半胱氨酸蛋白酶抑制剂有显著的同源性。重组蛋白在大肠杆菌中以GST融合的可溶性形式表达，并通过荧光底物分析鉴定其对木瓜蛋白酶、组织蛋白酶L和组织蛋白酶B的抑制活性。这是首次报道的细胞内半胱氨酸蛋白酶抑制剂同系物从蜱H。长角的(C)2008 Elsevier B. V.保留所有权利。
Cystatins are tight-binding inhibitors of papain-like cysteine proteases. On the basis of amino acid sequencing, cystatins can be subdivided into three closely related family 1, family 2 and family 3. Among them, only family I cystatins are intracellular. In this report, a gene encoding family 1 cystatin from tick Haemaphysalis longicornis (Hlcyst-1) has been identified. Its full-length cDNA is 437 bp, including an intact ORF encoding an expected protein with 98 amino acids. Sequence analysis demonstrated that it has significant homology with the known family 1 cystatin. The recombinant protein was expressed in a GST-fused soluble form in Escherichia coli, and its inhibitory activity against papain, cathepsin L, and cathepsin B was identified by fluorogenic substrate analysis. This is the first report of an intracellular cystatin homolog from the tick H. longicornis. (C) 2008 Elsevier B.V. All rights reserved.