Rhipicephalus sanguineus trypsin inhibitors present in the tick larvae:: isolation, characterization, and partial primary structure determination
Rhipicephalus sanguineus trypsin inhibitors present in the tick larvae:: isolation, characterization, and partial primary structure determination
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DOI：
10.1016/s0003-9861(03)00344-8
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发表时间：
2003-09-15
影响因子：
3.9
通讯作者：
Tanaka, AS
中科院分区：
文献类型：
--
作者：
Azzolini, SS;Sasaki, SD;Tanaka, AS
吸血动物是蛋白酶抑制剂的丰富来源，主要是那些干扰其宿主止血系统的蛋白酶抑制剂。蜱Rhipicephalus sanguineus是狗和其他动物的体外寄生虫。本工作的目的是纯化和表征丝氨酸蛋白酶抑制剂存在于R。血红幼虫（RsTI）。通过胰蛋白酶-琼脂糖凝胶亲和层析和Resource Q和Mono S柱离子交换层析分离抑制剂（RsTI）。当这些RsTI通过SDS-PAGE显示分子量在8和18 kDa之间时，它们在大约12个不同的蛋白峰中分离。纯化的RsTI对不同丝氨酸蛋白酶的特异性存在差异。对于中性粒细胞弹性蛋白酶、纤溶酶和HuPK，RsTIQ 2是比RsTIQ 7和RsTIS 5更好的抑制剂，解离常数（Ki）分别为1.3、3.2和22 nM。其他抑制剂如RsTIQ 7、RsTIS 3和RsTIS 5也影响嗜中性粒细胞弹性蛋白酶和纤溶酶，Ki在nM范围内。RsTIQ 2、RsTIQ 7和RsTIS 5氨基酸序列数据允许将它们分类为Kunitz型丝氨酸蛋白酶抑制剂家族的成员，尽管RsTI的作用仍然未知。我们的研究结果表明，丝氨酸蛋白酶抑制剂从R。血红牛蜱的抑制剂类似于来自微小牛蜱其他硬蜱物种的抑制剂，表明这些抑制剂在硬蜱物种中的类似作用，并且还作为产生或改进针对具有独特抗原的不同外寄生虫的疫苗的潜在工具。(C)2003年爱思唯尔公司All rights reserved.
Blood sucking animals are a rich source of proteinase inhibitors mainly those that interfere in their host hemostatic systems. The tick Rhipicephalus sanguineus is an ectoparasite of dogs and other animals. The aims of this work were the purification and characterization of serine proteinase inhibitors present in R. sanguineus larvae (RsTI). The inhibitors (RsTI) were isolated by affinity chromatography on trypsin-Sepharose and ion exchange chromatographies in Resource Q and Mono S columns. These RsTIs were separated in around 12 different protein peaks, when they showed molecular masses between 8 and 18 kDa, by SDS-PAGE. Purified RsTIs presented differences in the specificity for different serine proteinases. RsTIQ2 was, better inhibitor than RsTIQ7 and RsTIS5 for neutrophil elastase, plasmin, and HuPK with dissociation constants (K-i) of 1.3, 3.2, and 22 nM, respectively. Other inhibitors such as RsTIQ7, RsTIS3, and RsTIS5 also affected neutrophil elastase and plasmin with K-i in the nM range. The RsTIQ2, RsTIQ7, and RsTIS5 amino acid sequence data allowed classifying them as members of the Kunitz-type serine proteinase inhibitor family, even though the RsTI role is still unknown. Our present results showed that serine proteinase inhibitors from R. sanguineus are similar to inhibitors from Boophilus microplus other hard tick species, suggesting a similar role of these inhibitors in hard tick species and also as a potential tool to generate or improve vaccine against different ectoparasites with an unique antigen. (C) 2003 Elsevier Inc. All rights reserved.