定位于内质网膜的热休克蛋白90家族成员gp96蛋白作为一种对其“客户蛋白”具有高选择性的分子伴侣，与多种癌症的发生发展密切相关，而gp96-抗原肽复合物在癌症和传染性疾病的防治方向也已展现出广阔的应用前景。然而目前，领域内对于gp96与其辅助分子伴侣和“客户蛋白”(包括肿瘤抗原肽段)的互作机理， 以及gp96在ATP水解循环过程中的动态构象变化均不明晰。针对这一现状，本项目计划利用新兴的氢氘交换质谱技术手段并结合结构生物学的方法来研究上述科学问题。我们预期该研究成果将对gp96究竟如何利用ATP水解产生的能量来帮助其“客户蛋白”的正确折叠这个科学问题具有启示意义，同时也期待该研究成果能够对针对gp96的小分子或抗体抑制剂研发，以及gp96-抗原肽复合物疫苗进一步的改进和发展具有一定的指导意义。

As endoplasmic reticulum resident heat-shock protein 90 paralog, gp96 works as a highly selective molecular chaperone, whose client proteins embrace Toll like receptors (TLRs), HER2, integrins, etc. Despite the close relationship between up-regulation of gp96 and cancer growth/metastasis, and despite the therapeutic prospect of gp96-peptide complex in various cancers and infectious diseases, several fundamental biological questions of gp96 remained elusive for decades. These include the molecular basis of gp96’s high selectivity towards client proteins and its conformational dynamics along the ATP cycle. In light of this, we propose to use Hydrogen Deuterium Exchange coupled Mass Spectrometry (HDX-MS) together with structural biology and biochemical tools to elucidate gp96’s in solution conformational dynamics, and to reveal the molecular basis underlying its interaction with representative co-chaperon (PRAT4A) and client protein (TLRs). We anticipate that our work will shed light on the ultimate question: how the proper folding of client protein is achieved with the help from gp96 and its co-chaperon? And we hope that our work will contribute to the development of next generation gp96 inhibitors and benefit further clinical translation of gp96-peptide complex.