Mass spectrometric characterization of proteins from the SARS virus: a preliminary report.
Mass spectrometric characterization of proteins from the SARS virus: a preliminary report.
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DOI：
10.1074/mcp.m300048-mcp200
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发表时间：
2003-05
期刊：
影响因子：
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通讯作者：
Standing KG
中科院分区：
文献类型：
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作者：
Krokhin O;Li Y;Andonov A;Feldmann H;Flick R;Jones S;Stroeher U;Bastien N;Dasuri KV;Cheng K;Simonsen JN;Perreault H;Wilkins J;Ens W;Plummer F;Standing KG
一种新型冠状病毒被认为是严重急性呼吸系统综合征（SARS）的病原体。我们用几个SARS病人的恢复期血清检测另一个SARS病人灌洗细胞培养上清液中的蛋白质。通过基质辅助激光解吸/电离飞行时间质谱法（MALDI-TOF MS）鉴定上清液中最突出的蛋白质为约46-kDa物质。发现这是一种新的核衣壳蛋白，其与最近发表的同一病毒分离株的核苷酸序列中的开放阅读框所预测的几乎完全匹配（>96%覆盖率）。还通过MALDI-TOF MS（42%覆盖率）检查了与预测的139-kDa刺突糖蛋白对应的第二种病毒蛋白。经肽N-糖苷酶F消化后，确认了该蛋白中的12个糖基化位点。还鉴定了附着在其中四个位点上的糖。这些结果表明，核衣壳蛋白是一个主要的免疫原，可能是有用的早期诊断，刺突糖蛋白可能是一个特别有吸引力的目标，预防性干预，在打击SARS。
A new coronavirus has been implicated as the causative agent of severe acute respiratory syndrome (SARS). We have used convalescent sera from several SARS patients to detect proteins in the culture supernatants from cells exposed to lavage another SARS patient. The most prominent protein in the supernatant was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a ∼46-kDa species. This was found to be a novel nucleocapsid protein that matched almost exactly one predicted by an open reading frame in the recently published nucleotide sequence of the same virus isolate (>96% coverage). A second viral protein corresponding to the predicted ∼139-kDa spike glycoprotein has also been examined by MALDI-TOF MS (42% coverage). After peptide N-glycosidase F digestion, 12 glycosylation sites in this protein were confirmed. The sugars attached to four of the sites were also identified. These results suggest that the nucleocapsid protein is a major immunogen that may be useful for early diagnostics, and that the spike glycoprotein may present a particularly attractive target for prophylactic intervention in combating SARS.