Chemical probes to decipher PAD biology

项目摘要 蛋白质瓜氨酸在炎症性疾病和癌症中增加，证据表明， 翻译后修饰是疾病发病机制的主要贡献者。事实上，抑制蛋白质 瓜氨酸可降低类风湿性关节炎、溃疡性结肠炎、神经炎、 损伤和癌症。因此，催化该PTM的蛋白质精氨酸脱亚胺酶（PAD）代表了新的 治疗目标 本次更新申请的总体目标是研究瓜氨酸蛋白如何影响人类细胞 信号具体问题包括：（1）哪些蛋白质是瓜氨酸化的？(2)如何 瓜氨酸影响活性？(3)瓜氨酸会影响蛋白水解事件吗？（4）我们可以生成 同工酶特异性抑制剂？这些问题的答案将通过（1）发展化学蛋白质组学 （2）开发和使用遗传密码扩增平台， 评估个别瓜氨酸酶事件的功能后果;以及（3）开发化学探针 和同工酶选择性PAD抑制剂来调节细胞瓜氨酸水平。通过解决这些问题， 问题，PI希望提供关于PAD如何对人类生物学做出贡献的全面理解。 鉴于PI的出色记录及其在PAD领域的领导地位， 强大的合作者网络，该项目将提供新的见解和迫切需要的研究工具， 科学界。关键成果包括发现新的PAD生物学，预计将有 对自身免疫和癌症的主要翻译影响。

PROJECT SUMMARY Protein citrullination is increased in inflammatory disease and cancer, and the evidence indicates that this post-translational modification is a major contributor to disease pathogenesis. In fact, inhibition of protein citrullination decreases disease severity in animal models of rheumatoid arthritis, ulcerative colitis, nerve damage, and cancer. Thus, the Protein Arginine Deiminases (PADs), which catalyze this PTM, represent novel therapeutic targets. The overall goal of this renewal application is to investigate how protein citrullination impacts human cell signaling. Specific questions to be addressed include: (1) What proteins are citrullinated?; (2) How does citrullination affect activity?; (3) Does citrullination impact proteolytic events? and (4) Can we generate isozyme specific inhibitors? Answers to these questions will be obtained by (1) developing chemoproteomic platforms to identify citrullination events; (2) developing and using a genetic code expansion platform to evaluate the functional consequences of individual citrullination events; and (3) developing chemical probes and isozyme selective PAD inhibitors to modulate cellular citrullination levels. By addressing each of these questions, the PI expects to provide a holistic understanding for how the PADs contribute to human biology. Given the outstanding track record of the PI and his leadership position in the PAD field, combined with his strong network of collaborators, this project will provide new insights and sorely needed research tools to the scientific community. Key outcomes include the discovery of new PAD biology, which is predicted to have major translational impacts on autoimmunity and cancer.