Engineering protein-specific proteases: targeting signaling proteins

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.