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应用蛋白质组学分析技术对猪繁殖与呼吸综合征病毒粒子中的细胞蛋白进行分析。

Profiling of cellular proteins in porcine reproductive and respiratory syndrome virus virions by proteomics analysis.

机构信息

State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China.

出版信息

Virol J. 2010 Sep 18;7:242. doi: 10.1186/1743-422X-7-242.

Abstract

BACKGROUND

Porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped virus, bearing severe economic consequences to the swine industry worldwide. Previous studies on enveloped viruses have shown that many incorporated cellular proteins associated with the virion's membranes that might play important roles in viral infectivity. In this study, we sought to proteomically profile the cellular proteins incorporated into or associated with the virions of a highly virulent PRRSV strain GDBY1, and to provide foundation for further investigations on the roles of incorporated/associated cellular proteins on PRRSV's infectivity.

RESULTS

In our experiment, sixty one cellular proteins were identified in highly purified PRRSV virions by two-dimensional gel electrophoresis coupled with mass spectrometric approaches. The identified cellular proteins could be grouped into eight functional categories including cytoskeletal proteins, chaperones, macromolecular biosynthesis proteins, metabolism-associated proteins, calcium-dependent membrane-binding proteins and other functional proteins. Among the identified proteins, four have not yet been reported in other studied envelope viruses, namely, guanine nucleotide-binding proteins, tyrosine 3-monooxygenase/tryptophan 5-monooxygenase, peroxiredoxin 1 and galectin-1 protein. The presence of five selected cellular proteins (i.e., β-actin, Tubulin, Annexin A2, heat shock protein Hsp27, and calcium binding proteins S100) in the highly purified PRRSV virions was validated by Western blot and immunogold labeling assays.

CONCLUSIONS

Taken together, the present study has demonstrated the incorporation of cellular proteins in PRRSV virions, which provides valuable information for the further investigations for the effects of individual cellular proteins on the viral replication, assembly, and pathogenesis.

摘要

背景

猪繁殖与呼吸综合征病毒(PRRSV)是一种包膜病毒,给全球养猪业带来了严重的经济后果。先前对包膜病毒的研究表明,许多与病毒包膜相关的细胞蛋白可能在病毒感染性中发挥重要作用。在这项研究中,我们试图对高度传染性 PRRSV 株 GDBY1 的病毒粒子中掺入或与之相关的细胞蛋白进行蛋白质组学分析,并为进一步研究掺入/相关细胞蛋白对 PRRSV 感染性的作用提供基础。

结果

在我们的实验中,通过二维凝胶电泳结合质谱方法,在高度纯化的 PRRSV 病毒粒子中鉴定出 61 种细胞蛋白。鉴定出的细胞蛋白可分为八个功能类别,包括细胞骨架蛋白、伴侣蛋白、大分子生物合成蛋白、代谢相关蛋白、钙依赖性膜结合蛋白和其他功能蛋白。在鉴定出的蛋白质中,有四种尚未在其他研究的包膜病毒中报道过,即鸟嘌呤核苷酸结合蛋白、酪氨酸 3-单加氧酶/色氨酸 5-单加氧酶、过氧化物酶 1 和半乳糖凝集素-1 蛋白。Western blot 和免疫金标记检测验证了β-肌动蛋白、微管蛋白、膜联蛋白 A2、热休克蛋白 Hsp27 和钙结合蛋白 S100 等五种选定的细胞蛋白在高度纯化的 PRRSV 病毒粒子中的存在。

结论

综上所述,本研究表明细胞蛋白被掺入 PRRSV 病毒粒子中,这为进一步研究单个细胞蛋白对病毒复制、组装和发病机制的影响提供了有价值的信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/80e2/2949843/9eb7a45475a1/1743-422X-7-242-1.jpg

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