Resch Wolfgang, Hixson Kim K, Moore Ronald J, Lipton Mary S, Moss Bernard
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 4 Center Drive, MSC 0445, Bethesda, MD 20892-0445, USA.
Virology. 2007 Feb 5;358(1):233-47. doi: 10.1016/j.virol.2006.08.025. Epub 2006 Sep 26.
The protein content of vaccinia virus mature virions, purified by rate zonal and isopycnic centrifugations and solubilized by SDS or a solution of urea and thiourea, was determined by the accurate mass and time tag technology which uses both tandem mass spectrometry and Fourier transform-ion cyclotron resonance mass spectrometry to detect tryptic peptides separated by high-resolution liquid chromatography. Eighty vaccinia virus-encoded proteins representing 37% of the 218 genes annotated in the complete genome sequence were detected in at least three analyses. Ten proteins accounted for approximately 80% of the virion mass. Thirteen identified proteins were not previously reported as components of virions. On the other hand, 8 previously described virion proteins were not detected here, presumably due to technical reasons including small size and hydrophobicity. In addition to vaccinia virus-encoded proteins, 24 host proteins omitting isoforms were detected. The most abundant of these were cytoskeletal proteins, heat shock proteins and proteins involved in translation.
通过速率区带离心和等密度离心纯化、并用十二烷基硫酸钠(SDS)或尿素与硫脲溶液溶解的痘苗病毒成熟病毒粒子的蛋白质含量,采用精确质量和时间标签技术进行测定,该技术利用串联质谱和傅里叶变换离子回旋共振质谱来检测通过高分辨率液相色谱分离的胰蛋白酶肽段。在至少三次分析中检测到了80种痘苗病毒编码的蛋白质,占完整基因组序列中注释的218个基因的37%。10种蛋白质约占病毒粒子质量的80%。13种已鉴定的蛋白质以前未被报道为病毒粒子的成分。另一方面,这里未检测到8种先前描述的病毒粒子蛋白质,可能是由于包括体积小和疏水性在内的技术原因。除了痘苗病毒编码的蛋白质外,还检测到24种宿主蛋白质(不包括异构体)。其中最丰富的是细胞骨架蛋白、热休克蛋白和参与翻译的蛋白质。