Engineering protein-specific proteases: targeting signaling proteins

项目摘要 这一补充申请的目标是成功地重新整合博士贝蒂朱进入研究 社区在第3年和第4年的父R 01奖。朱博士是一位非常有前途的结构生物学家， 于2020年5月获得X射线晶体学领域的博士学位，该领域应用于新的，非 典型的DNA结构她在高影响力期刊（Journal）上获得了两篇第一作者同行评审论文 美国化学学会和核酸研究）从她的论文研究，以及一个 奖项的数量。她的计划是在她最初的博士后研究中将她的技能扩展到蛋白质结构的研究 预约然而，由于恶劣的研究环境对她的心理健康产生了负面影响， 受到了精神创伤，被迫离开了这个职位。经过大约18个月的中断，她在 在过去的三个月里，奥班/托特实验室作为她康复的一部分，并为她测试沃茨。 完全重新融入生物医学研究界。附加奖是一个优秀的 机制，以帮助朱博士完全重新融入生物医学研究界，为她提供 独立的支持，使她能够继续追求她的职业目标。 我们的辅导计划符合朱博士的职业目标，并允许她恢复研究 在一个安全，支持和培育的环境中对蛋白质结构生物学感兴趣。指导计划是 分为两个阶段。在第一阶段，朱博士将与托特博士在X射线结构方面密切合作 确定进化的蛋白酶及其与肽的复合物，这是研究计划的一个方面， 比较简单，支持母奖项的目标1。这会让朱博士重新认识 结构生物学研究环境，使她重新获得信心，同时获得新的技术技能。 我们预计，该计划的这一阶段将需要大约6至9个月的时间。在第二阶段，博士。 朱棣文将开始承担资助研究计划中更雄心勃勃的方面，特别是X射线 设计的蛋白酶和全长RAS之间的复合物的晶体结构测定 这也支持父母补助金的目标1。此外，她将在此阶段获得新的细胞生物学技能， 在真核细胞模型系统中测试有前途的蛋白酶，这将为结构-功能 作为母提案中目标3的一部分。第二阶段将是最后的十五年， 18个月的补充。 我们预计，朱博士将在一个位置，以申请F32博士后奖学金，在她的 第二年，让她回到正轨，追求她的目标，一个独立的研究生涯。我们认为 她在计划外中断科学研究之前的记录是她具有巨大潜力的有力证据 在适当的指导下做出重要的研究贡献。

Project Summary The goal of this supplement application is to successfully re-integrate Dr. Betty Chu into the research community over Years 3 and 4 of the parent R01 award. Dr. Chu is a very promising structural biologist who received her PhD in May 2020 in the field of X-ray crystallography as applied to the study of novel, non- canonical DNA structures. She obtained two first author peer-reviewed papers in high impact journals (Journal of the American Chemical Society and Nucleic Acids Research) from her dissertation research, as well as a number of awards. Her plan was to extend her skills to the study of protein structure in her initial postdoctoral appointment. However, due to a hostile research environment that negatively impacted her mental health, she was traumatized and forced to leave that position. After a hiatus of approximately 18 months, she worked in the Orban/Toth laboratory for the past three months as part of her recovery, and to test the waters for her complete re-integration into the biomedical research community. The supplemental award is an excellent mechanism to help Dr. Chu fully re-integrate into the biomedical research community by providing her with independent support so that she can resume the pursuit of her career goals. Our mentoring plan is in line with Dr. Chu's career goals and allows her to resume her research interests in protein structural biology in a safe, supportive and nurturing environment. The mentoring plan is organized into two phases. In the first phase, Dr. Chu will work closely with Dr. Toth on X-ray structure determination of evolved proteases and their complexes with peptides, an aspect of the research program that is comparatively straightforward and supports Aim 1 of the parent award. This will re-introduce Dr. Chu to a structural biology research setting and enable her to regain her confidence while acquiring new technical skills. We anticipate that this phase of the plan will take approximately six to nine months. In the second phase, Dr. Chu will begin to undertake more ambitious aspects of the funded research program, in particular, X-ray crystallographic structure determination of the complexes between designed proteases and full-length RAS that also supports Aim 1 of the parent grant. Additionally, she will acquire new cell biology skills in this phase, testing promising proteases in eukaryotic cell model systems, which will provide insight into structure-function relationships as part of Aim 3 in the parent proposal. The second phase will constitute the final fifteen to eighteen months of the supplement. We anticipate that Dr. Chu will be in a position to apply for an F32 Postdoctoral Fellowship in her second year, putting her back on track to pursue her objective of an independent research career. We believe her track record prior to her unplanned hiatus from science is strong evidence that she has significant potential to make important research contributions with proper mentoring.