Chemical Approaches for Activity-Based Proteomics

根据活性表征整个蛋白质类别的能力将极大地加速 蛋白质在基因组时代的功能定位及新生物标记物和治疗药物的鉴定 人类疾病诊断和治疗的目标。本着这一目标，我们创造了创新 化学蛋白质组平台，用于描述天然生物系统中的整体蛋白质活性。在过去的拨款中 在此期间，我们提出并应用了这些平台，取得了以下主要成果：1) 发现和抑制去调节的、促肿瘤的脂水解酶；2)开发一种 蛋白质组平台，发现新形式的半胱氨酸天冬氨酸氨基转移酶-激酶串扰，调节肿瘤细胞凋亡；3) 发现一种新的翻译后修饰，将葡萄糖摄取与糖酵解重塑和 促肿瘤代谢物的生产；4)基于无标签活性的蛋白质图谱的建立 (ABPP)平台，用于量化和功能表征癌症中的反应性半胱氨酸；以及5)进展 无标签ABPP以优化作为抗癌药物的共价激酶抑制剂的活性和选择性。在……里面 这次续期申请，我们将在以往成果的基础上，检验以下主要假设 对癌症和化学生物学的高度意义：1)脂水解酶是新陈代谢之间的关键桥梁 和促进肿瘤发生的信号通路；2)蛋白分解和磷酸化之间的串扰 途径调节癌细胞存活并产生肿瘤特异性生物标记物用于监测癌症治疗 和抗性；3)3-磷酸甘油-赖氨酸(PGK)修饰构成了一条内在的反馈途径 重组糖酵解产物并刺激肿瘤生长；4)电泳性/氧化敏感的半胱氨酸调节 癌细胞对氧化应激反应的存活；以及5)无标签优化的共价激酶抑制剂 ABPP将显示增强的目标上的活动和减少的目标外交互，从而扩展其 作为治疗窗口的抗癌药物。这个应用程序的最终目标是识别关键的生化 支持人类癌症生长和恶性肿瘤的途径。这些途径的分子组成 可能代表癌症的新生物标记物和药物靶点。

The ability to characterize entire classes of proteins based on activity would greatly accelerate both the assignment of protein function in the genome era and the identification of new biomarl<ers and therapeutic targets for the diagnosis and treatment of human disease. With this goal in mind, we have created innovative chemoproteomic platforms to profile protein activities en masse in native biological systems. In the past grant period, we advanced and applied these platforms and the following major achievements were attained: 1) discovery and inhibition of deregulated, pro-tumorigenic lipid hydrolases; 2) development of an integrated proteomic platform to discover novel forms of caspase-kinase crosstalk that regulate cancer apoptosis; 3) discovery of a novel post-translational modification that couples glucose uptake to glycolytic remodeling and the production of pro-tumorigenic metabolites; 4) development of a tag-free activity-based protein profiling (ABPP) platform to quantify and functionally characterize reactive cysteines in cancer; and 5) advancement of tag-free ABPP to optimize the activity and selectivity covalent kinase inhibitors as anti-cancer drugs. In this renewal application, we will build upon our past achievements to test the following major hypotheses of high significance to cancer and chemical biology: 1) lipid hydrolases form key bridges between metabolic and signaling pathways that promote tumorigenesis; 2) crosstalk between proteolytic and phosphorylation pathways regulate cancer cell survival and create tumor-specific biomarkers for monitoring cancer therapy and resistance; 3) 3-phosphoglyceryl-lysine (pgK) modifications constitute an intrinsic feedback pathway to remodel glycolytic output and stimulate tumor growth; 4) electrophile/oxidation-sensitive cysteines regulate cancer cell survival in response to oxidative stress; and 5) covalent kinase inhibitors optimized by tag-free ABPP will show enhanced on-target activity and reduced off-target interactions, thereby expanding their therapeutic window as anti-cancer drugs. The ultimate goal ofthis application is to Identify key biochemical pathways that support human cancer growth and malignancy. The molecular components of these pathways may represent new biomarkers and drug targets for cancer.