Proteomic identification of proteins conjugated to ISG15 in mouse and human cells
Proteomic identification of proteins conjugated to ISG15 in mouse and human cells
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DOI：
10.1016/j.bbrc.2005.08.132
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发表时间：
2005-10-21
影响因子：
3.1
通讯作者：
Zhang, DE
中科院分区：
文献类型：
--
作者：
Giannakopoulos, NV;Luo, JK;Zhang, DE
虽然干扰素诱导蛋白ISG 15是最早被发现的泛素样修饰物之一，但关于与ISG 15缀合的蛋白质的身份或修饰的生物学后果仍有许多未知之处。为了更好地了解受ISG 15影响的细胞途径，我们使用蛋白质组学方法鉴定了ISGylation靶向蛋白。质谱分析鉴定了来自干扰素处理的小鼠或人细胞的抗ISG 15免疫沉淀物中的76个候选ISGylation靶标。在小鼠和人类样本中发现了21种蛋白质，包括STAT 1，一种已知的ISGylation靶标。在两个物种中鉴定的候选物在转染系统中测试ISG化：19种测试的蛋白质中的18种在该系统中被ISG化。两个候选物，EF-2和VCP，也显示在不存在外源性过表达的情况下以干扰素依赖性方式ISGylated。从单个物种中鉴定出的七种蛋白质，但功能上与两个物种中发现的候选蛋白相关，也在过表达系统中被ISGylated。可被ISG化的蛋白质在翻译、糖酵解、应激反应和细胞运动中起重要作用。这些数据表明ISGylation靶向在几个基本重要的细胞途径中发现的蛋白质，并将有助于理解干扰素诱导的ISG 15和ISG 15缀合的生理作用。(c)2005年爱思唯尔公司All rights reserved.
Though the interferon-inducible protein ISG15 was one of the first ubiquitin-like modifiers to be discovered, much remains unknown about the identity of proteins conjugated to ISG15 or the biologic consequences of modification. To gain a better understanding of the cellular pathways affected by ISG15, we identified proteins targeted for ISGylation using a proteomic approach. Mass spectrometric analysis identified 76 candidate ISGylation targets in anti-ISG15 immunoprecipitates from interferon-treated mouse or human cells. Twenty-one proteins were found in both mouse and human samples, including STAT1, a known target of ISGylation. Candidates identified in both species were tested for ISGylation in a transfection system: 18 of 19 proteins tested were ISGylated in this system. Two candidates, EF-2 and VCP, were also shown to be ISGylated in an interferon-dependent manner in the absence of exogenous over-expression. Seven proteins identified from a single species, but functionally related to candidates found in both species, were also ISGylated in the over-expression system. Proteins that can be ISGylated play important roles in translation, glycolysis, stress responses, and cell motility. These data indicate that ISGylation targets proteins found in several fundamentally important cellular pathways and will contribute to understanding the physiologic role of interferon-induced ISG15 and ISG15 conjugation. (c) 2005 Elsevier Inc. All rights reserved.