ORGANELLAR PROTEOMICS REVEALS GOLGI ARGININE DIMETHYLATION

细胞器蛋白质组学揭示高尔基精氨酸二甲基化

• 批准号: 7420652
• 负责人: KATHRYN Elizabeth HOWELL
• 金额: $ 0.29万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-20 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7420652
• 关键词: arginine; proteins

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Golgi complex functions to posttranslationally modify newly synthesized proteins and lipids and to sort them to their sites of function. In this study, a stacked Golgi fraction was isolated by classical cell fractionation, and the protein complement (the Golgi proteome) was characterized using multidimensional protein identification technology. Many of the proteins identified are known residents of the Golgi, and 64% of these are predicted transmembrane proteins. Proteins localized to other organelles also were identified, strengthening reports of functional interfacing between the Golgi and the endoplasmic reticulum and cytoskeleton. Importantly, 41 proteins of unknown function were identified. Two were selected for further analysis, and Golgi localization was confirmed. One of these, a putative methyltransferase, was shown to be arginine dimethylated, and upon further proteomic analysis, arginine dimethylation was identified on 18 total proteins in the Golgi proteome. This survey illustrates the utility of proteomics in the discovery of novel organellar functions and resulted in 1) a protein profile of an enriched Golgi fraction; 2) identification of 41 previously uncharacterized proteins, two with confirmed Golgi localization; 3) the identification of arginine dimethylated residues in Golgi proteins; and 4) a confirmation of methyltransferase activity within the Golgi fraction.

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。高尔基复合体的功能是翻译后修饰新合成的蛋白质和脂质，并将它们分类到它们的功能位点。本研究采用经典细胞分离技术分离堆叠高尔基体片段，并利用多维蛋白鉴定技术对蛋白质补体（高尔基蛋白组）进行表征。许多被鉴定的蛋白质是已知的高尔基体的居民，其中64%是预测的跨膜蛋白质。定位于其他细胞器的蛋白质也被发现，加强了高尔基体与内质网和细胞骨架之间功能界面的报道。重要的是，鉴定了41种功能未知的蛋白质。其中2例进行进一步分析，确认高尔基定位。其中一个假定的甲基转移酶被证明是精氨酸二甲基化的，在进一步的蛋白质组学分析中，精氨酸二甲基化在高尔基蛋白组中的18个蛋白上被鉴定出来。这项调查说明了蛋白质组学在发现新的细胞器功能方面的效用，并导致1)富含高尔基部分的蛋白质谱；2)鉴定了41个先前未被鉴定的蛋白，其中2个已确定高尔基定位；3)高尔基蛋白精氨酸二甲基化残基的鉴定；4)确认高尔基体片段内的甲基转移酶活性。