MAPPING PHOSPHORYLATION SITES IN THE NON-STRUCTURAL PROTEIN 5A FROM HEPATITUS C

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。丙型肝炎病毒编码一种被称为NS5a的定义不明确的蛋白。据报道，NS5a被磷酸化，但修饰的氨基酸尚不清楚。Rice实验室的Evans博士为我们提供了HCV复制子感染肝细胞的NS5a亲和纯化版本。我们使用假设驱动质谱法寻找并鉴定NS5a磷酸化位点。到目前为止，我们已经确定了NS5a上的一个新的磷酸化位点。该项目目前的工作包括：1)突变磷酸化位点并确定对病毒的影响2)修改NS5a的纯化技术，以便对相互作用蛋白进行纯化和质谱鉴定。

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 hepatitis C virus encodes a poorly defined protein termed NS5a. NS5a has been reported to be phosphorylated, but the amino acids modified are not known. Dr. Evans from the Rice laboratory has provided us with an affinity purified version of NS5a from HCV replicon-infected hepatocytes. We used hypothesis-driven mass spectrometry to search for and identify sites of phosphorylation on NS5a. To date, we have identified one novel site of phosphorylation on NS5a. Current efforts on this project: 1) mutate the site of phosphorylation and determine effects on the virus 2) modify the purification techniques for NS5a to allow for purification and mass spectrometric identification of interacting proteins.