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一种蛋白质组学方法,用于分析软蜱 Ornithodoros savignyi 在受到白色念珠菌挑战时参与即时防御反应的血淋巴蛋白。

A proteomics approach for the analysis of hemolymph proteins involved in the immediate defense response of the soft tick, Ornithodoros savignyi, when challenged with Candida albicans.

机构信息

Department of Biochemistry, University of Pretoria, South Africa.

出版信息

Exp Appl Acarol. 2010 Aug;51(4):309-25. doi: 10.1007/s10493-010-9338-z. Epub 2010 Feb 26.

DOI:10.1007/s10493-010-9338-z
PMID:20186467
Abstract

A proteomics approach was employed to identify proteins secreted into the hemolymph of Ornithodorus savignyi ticks 2 h after immune-challenge with the yeast, Candida albicans. Profiling of the proteins present in hemolymph of unchallenged ticks versus ticks challenged with heat-killed yeast revealed five proteins to be differentially expressed. The modulated protein spots were subjected to tandem mass spectrometry (MS/MS) analysis, but could not be positively identified. These proteins can be assigned to the immune response as they were not induced after aseptic injury. In an attempt to identify hemolymph proteins that recognize and bind to yeast cells, hemolymph obtained from both unchallenged and challenged ticks was incubated with C. albicans. Elution of the bound proteins followed by SDS-PAGE analysis indicated that three proteins (97, 88 and 26 kDa) present in both unchallenged and challenged hemolymph samples bind to yeast cells. The constant presence of these three proteins in tick hemolymph leads us to believe that they may be involved in non-self recognition and participate in yeast clearance from tick plasma. The analyzed yeast-binding proteins could also not be positively identified, suggesting that all the tick immune proteins investigated in this study are novel.

摘要

采用蛋白质组学方法鉴定了在感染白念珠菌酵母 2 小时后,革螨 Ornithodorus savignyi 血淋巴中分泌的蛋白质。与热杀死酵母刺激的革螨相比,未刺激革螨的血淋巴中存在的蛋白质进行了分析,发现有 5 种蛋白质表达水平存在差异。对这些被调控的蛋白斑点进行串联质谱(MS/MS)分析,但无法被明确鉴定。这些蛋白质可被分配到免疫反应中,因为它们不会在无菌损伤后被诱导。为了鉴定识别并结合酵母细胞的血淋巴蛋白,从未刺激和刺激的革螨中获得的血淋巴与白念珠菌孵育。结合蛋白的洗脱后进行 SDS-PAGE 分析表明,在未刺激和刺激的血淋巴样品中存在三种结合到酵母细胞上的蛋白(97、88 和 26 kDa)。这三种蛋白在革螨血淋巴中的持续存在使我们相信它们可能参与非自我识别,并参与从革螨血浆中清除酵母。分析的酵母结合蛋白也无法被明确鉴定,这表明本研究中所有被检测的革螨免疫蛋白均为新型蛋白。

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