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微小隐孢子虫侵袭蛋白的鉴定

Identification of invasion proteins of Cryptosporidium parvum.

作者信息

Singh Preeti, Mirdha Bijay Ranjan, Srinivasan Alagiri, Rukmangadachar Lokesh A, Singh Sundeep, Sharma Pratibha, Hariprasad Gururao, Gururao Hariprasad, Luthra Kalpana

出版信息

World J Microbiol Biotechnol. 2015 Dec;31(12):1923-34. doi: 10.1007/s11274-015-1936-9.

Abstract

Host cell interactions and invasion by Cryptosporidium is a complex process mediated by zoites ligand-host cell receptors. Knowledge of proteins involved in this process will enable entry level inhibitors to be tried as therapeutic agents. In the present study, invasion proteins of Cryptosporidium parvum were studied in vitro. Cryptosporidium sporozoites membrane proteins were isolated and Cy5 dye labelled. They were then allowed to interact with the intact host cells. The interacting proteins were identified using 2-dimensional gel electrophoresis followed by mass spectrometry analysis. Sixty-one proteins were identified including twenty-seven previously reported invasion proteins. The newly identified proteins such as serine/threonine protein kinase, PI4 kinase, Hsp105 and coiled coil may have their roles in the parasitic invasion process. Thus, a new approach was used in the study to identify the probable proteins involved in invasion and/or host-parasite interactions. The advantage of this method is that it takes only a months' time instead of decades to identify these proteins involved in invasion process.

摘要

隐孢子虫与宿主细胞的相互作用及入侵是一个由子孢子配体-宿主细胞受体介导的复杂过程。了解参与这一过程的蛋白质将有助于尝试将入门级抑制剂用作治疗药物。在本研究中,对微小隐孢子虫的入侵蛋白进行了体外研究。分离出隐孢子虫子孢子膜蛋白并用Cy5染料标记。然后让它们与完整的宿主细胞相互作用。使用二维凝胶电泳随后进行质谱分析来鉴定相互作用的蛋白质。鉴定出了61种蛋白质,其中包括27种先前报道的入侵蛋白。新鉴定出的蛋白质,如丝氨酸/苏氨酸蛋白激酶、PI4激酶、热休克蛋白105和卷曲螺旋结构,可能在寄生虫入侵过程中发挥作用。因此,本研究采用了一种新方法来鉴定可能参与入侵和/或宿主-寄生虫相互作用的蛋白质。这种方法的优点是识别参与入侵过程的这些蛋白质只需要一个月的时间,而不是几十年。

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