University of North Carolina, Chapel Hill, NC, United States.
Department of Molecular Biology and Biochemistry, UC-Irvine, Irvine, CA 92697, United States.
Virology. 2014 Mar;452-453:310-23. doi: 10.1016/j.virol.2014.01.012. Epub 2014 Feb 25.
To the best of our knowledge, two phosphorylation sites have been reported previously, among 11 known Vaccinia virus phosphoproteins. Here, via phosphopeptide mass spectrometry, up to 189 phosphorylation sites were identified among 48 proteins in preparations of purified Vaccinia mature virus (MV). 8.5% of phospho-residues were pTyr. Viral phosphoproteins were found in diverse functional classes, including structural proteins, membrane proteins and RNA polymerase subunits. Among the nine identified membrane phosphoproteins, the sites in just one, namely A14L, were deduced to be internal with respect to the accompanying membrane. Examination of sites in known substrates of the Vaccinia-encoded protein kinase VPK2, indicated VPK2 to be a proline-dependent kinase. The MV phosphoproteome was enriched in potential substrates of cellular kinases belonging to the CDK2/CDK3, CK2, and p38 groups. Quantitative mass spectrometry identified several sites that became phosphorylated during intravirion kinase activation in vitro, each showing one of two distinct pH-dependency profiles.
据我们所知，在 11 种已知的痘苗病毒磷酸化蛋白中，以前已经报道了两个磷酸化位点。在这里，通过磷酸肽质谱分析，在纯化的痘苗成熟病毒 (MV) 制剂中 48 种蛋白质中鉴定出多达 189 个磷酸化位点。磷酸化残基的 8.5%为 pTyr。病毒磷酸化蛋白存在于多种功能类别中，包括结构蛋白、膜蛋白和 RNA 聚合酶亚基。在鉴定出的 9 种膜磷酸化蛋白中，只有一个位点（即 A14L）被推断为与其伴随的膜内部。对痘苗病毒编码的蛋白激酶 VPK2 的已知底物中的位点进行检查表明，VPK2 是一种依赖脯氨酸的激酶。MV 磷酸化组富含属于 CDK2/CDK3、CK2 和 p38 组的细胞激酶的潜在底物。定量质谱分析鉴定出在体外病毒激酶激活过程中发生磷酸化的几个位点，每个位点都显示出两种不同的 pH 依赖性谱之一。
