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刚果锥虫昆虫期表面抗原(TcCISSA)的纯化、结晶及X射线衍射分析

Purification, crystallization and X-ray diffraction analysis of Trypanosoma congolense insect-stage surface antigen (TcCISSA).

作者信息

Tonkin Michelle L, Workman Sean D, Eyford Brett A, Loveless Bianca C, Fudge Jessica L, Pearson Terry W, Boulanger Martin J

机构信息

Department of Biochemistry and Microbiology, University of Victoria, Petch Building, 3800 Finnerty Road, Victoria, BC V8P 5C2, Canada.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Dec 1;68(Pt 12):1503-6. doi: 10.1107/S1744309112042686. Epub 2012 Nov 14.

Abstract

Trypanosoma congolense is a major contributor to the vast socioeconomic devastation in sub-Saharan Africa caused by animal African trypanosomiasis. These protozoan parasites are transmitted between mammalian hosts by tsetse-fly vectors. A lack of understanding of the molecular basis of tsetse-trypanosome interactions stands as a barrier to the development of improved control strategies. Recently, a stage-specific T. congolense protein, T. congolense insect-stage surface antigen (TcCISSA), was identified that shows considerable sequence identity (>60%) to a previously identified T. brucei insect-stage surface molecule that plays a role in the maturation of infections. TcCISSA has multiple di-amino-acid and tri-amino-acid repeats in its extracellular domain, making it an especially interesting structure-function target. The predicted mature extracellular domain of TcCISSA was produced by recombinant DNA techniques, purified from Escherichia coli, crystallized and subjected to X-ray diffraction analysis; the data were processed to 2.7 Å resolution.

摘要

刚果锥虫是由动物非洲锥虫病导致撒哈拉以南非洲巨大社会经济破坏的主要原因之一。这些原生动物寄生虫通过采采蝇媒介在哺乳动物宿主之间传播。对采采蝇 - 锥虫相互作用分子基础的缺乏了解,是阻碍改进控制策略发展的一个障碍。最近,一种阶段特异性的刚果锥虫蛋白,即刚果锥虫昆虫阶段表面抗原(TcCISSA)被鉴定出来,它与先前鉴定的在感染成熟过程中起作用的布氏锥虫昆虫阶段表面分子具有相当高的序列同一性(>60%)。TcCISSA在其细胞外结构域中有多个二氨基酸和三氨基酸重复序列,使其成为一个特别有趣的结构 - 功能靶点。通过重组DNA技术产生了TcCISSA预测的成熟细胞外结构域,从大肠杆菌中纯化出来,进行结晶并进行X射线衍射分析;数据处理后分辨率达到2.7 Å。

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