A mass spectrometry-based proteornic approach to study Marek's Disease Virus gene expression
A mass spectrometry-based proteornic approach to study Marek's Disease Virus gene expression
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DOI：
10.1016/j.jviromet.2006.02.001
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发表时间：
2006-07-01
影响因子：
3.1
通讯作者：
Goshe, Michael B.
中科院分区：
文献类型：
--
作者：
Liu, Hsiao-Ching S.;Soderblom, Erik J.;Goshe, Michael B.
马立克氏病病毒（MDV）是一种鸟类疱疹病毒，可引起鸡的淋巴组织增生性疾病。MDV在产生新病毒的裂解期和病毒休眠的潜伏期之间转换。控制这种溶解性到潜伏性转换的机制仍不清楚。为了更好地了解MDV感染的裂解期，开发了基于质谱的策略来鉴定病毒蛋白并定性检查其在裂解感染的鸡胚成纤维细胞（CEF）中的丰度。强阳离子交换色谱（SCXC）和微毛细管反相液相色谱-串联质谱（mu rpLC/MS/MS）的组合用于解析来自MDV感染的CEF细胞裂解物的胰蛋白酶解物的肽。通过使用SEQUEST算法针对包含MDV蛋白和所有当前可用的原鸡蛋白的蛋白质数据库搜索串联质谱来鉴定肽。共鉴定出427种MDV肽，对应于82种独特蛋白，其中56种至少检测到两种独特肽。总体而言，近80%的所有推定的MDV蛋白表达感染的CEF细胞被确定。我们预计，这种方法将是一种可行的方法，用于确定病毒和宿主蛋白质组的变化发生在马立克氏病的发病机制，调节之间的切换溶解和潜伏期的MDV生命周期。(c)2006 Elsevier B. V.保留所有权利。
Marek's Disease Virus (MDV) is an avian herpesvirus that causes a lymphoproliferative disorder in chickens. MDV transitions between a lytic phase in which new viruses are produced and a latent phase in which the virus lays dormant. The mechanism controlling this lytic-to-latent switch remains unclear. To better understand the lytic phase of MDV infection, a mass spectrometry-based strategy was developed to identify viral proteins and to qualitatively examine their abundance in lytically infected chicken embryo fibroblast (CEF) cells. A combination of strong cation exchange chromatography (SCXC) and microcapillary reversed-phase liquid chromatography-tandem mass spectrometry (mu rpLC/MS/MS) was used to resolve peptides from tryptic digests of MDV-infected CEF cell lysates. Peptides were identified by searching the tandem mass spectra against a protein database containing both MDV proteins and all currently available Gallus gallus proteins using the SEQUEST algorithm. A total of 427 MDV peptides, corresponding to 82 unique proteins, were identified, with 56 of them detected with at least two unique peptides. Overall, nearly 80% of all putative MDV proteins expressed in infected CEF cells were identified. We anticipate that this approach will be a viable method for determining how viral and host proteome changes occurring in Marek's Disease pathogenesis regulate the switch between the lytic and latent phases of the MDV life cycle. (c) 2006 Elsevier B.V. All rights reserved.