CAREER: Using chemistry to probe anthrax toxin protein translocation

麻省理工学院的布拉德利·L·彭特鲁特将获得这一奖项，该奖项由生命过程化学计划资助，他将研究蛋白质跨细胞膜转移的机制。自然界进化出了许多大的蛋白质，它们的功能是进入哺乳动物细胞。其中一些蛋白质是由细菌产生的，通常对宿主有毒。炭疽毒素就是这种蛋白质的一个例子。这项研究的目的是用化学方法揭示炭疽毒素蛋白进入细胞的分子基础。一组易位分析和一个允许随意修改的蛋白质修改工具包将用于研究无毒形式的炭疽毒素是如何进入细胞的。通过采用化学方法，将制备和研究在这些大型炭疽蛋白质复合体中的特定位置包含化学修饰的变体，以便提供对这些纳米机器操作的基本了解。研究生和本科生的研究培训将使学生接触到这些研究中使用的最新化学方法，并为他们提供对复杂生物过程的洞察。更多的努力将被投入到一个外展的化学品展览中。PI将与麻省理工学院的本科生一起表演对8-12岁的学生来说似乎具有魔力的化学演示。魔术的教学理念是吸引年轻人的注意力，激发他们对科学的兴趣。推广计划的长期目标是引导年轻的赞助人进入基础科学的职业生涯。这项研究项目将系统地揭示Cargo在跨膜转运中的作用，并设计带电多肽，使异源蛋白通过保护性的抗原孔。研究程序将使用蛋白质修改工具包，允许随意修改和研究炭疽毒素纳米机器。本文提出的研究将提供对蛋白质跨膜转运的机制的理解，并为使用半合成策略来探索蛋白质转运过程奠定基础。此外，从这些研究中收集到的工具和见解将允许开发将新的化学形式输送到细胞中的抑制剂和方法，这是化学中的一个长期问题。

With this award, Bradley L. Pentelute from the Massachusetts Institute of Technology, funded by the Chemistry of Life Processes Program, will investigate the mechanism of protein translocation across a cell membrane. Nature has evolved a number of large proteins whose function is to enter mammalian cells. Some of these proteins are produced by bacteria and are often toxic to the host. An example of such a protein is anthrax toxin. The aim of the research is to use chemistry to uncover the molecular basis of anthrax toxin protein entry into cells. A battery of translocation assays and a protein modification toolkit, that allows at-will modification, will be used to investigate how the non-toxic forms of anthrax toxin enter cells. By virtue of taking a chemical approach, variants that contain chemical modifications at defined positions within these large anthrax protein complexes will be prepared and investigated in order to provide a fundamental understanding of the operation of these nanomachines. Research training at the graduate and undergraduate levels will expose students to the latest chemical methods that are used in these studies and provide them with insight into complicated biological processes. Additional effort will be put into an outreach chemical show. The PI together with MIT undergraduate students will perform chemical demonstrations that appear to be magical to an audience of 8-12 year old students. The teaching philosophy is that magic will capture the youths' attention and excite them about science. The long-term aim of outreach program is to springboard young patrons into careers of fundamental science.This research project will systematically uncover the role of cargo on translocation across a membrane and design charged peptides for heterologous protein passage through a protective antigen pore. The research procedures will employ a protein modification toolkit that allows at-will modification and investigation of the anthrax toxin nanomachine. The studies proposed here will provide a mechanistic understanding of protein translocation across a membrane and lay the foundation for using semi-synthetic strategies to probe the translocation process. In addition, the tools and insights gleaned from these studies will allow the development of inhibitors and methods to deliver novel chemical forms into cells, which is a longstanding problem in chemistry.