# Chemical Approaches for Interrogating Fundamental Biomedical Processes

> **NIH NIH R35** · UNIVERSITY OF VIRGINIA · 2020 · $343,392

## Abstract

Project Summary
 The development and application of novel chemical tools can provide fundamental insights into
biomedical processes. The research program outlined in this proposal focuses on three core areas: 1) the
development and application of chemosensors for phospho-regulatory enzymes, 2) new methodologies for
constructing designer signaling networks, and 3) fundamental studies of protein misfolding.
 Protein phosphorylation plays a central role in cellular signaling. Consequently, the enzymes that
maintain the phosphoproteome, protein kinases and protein phosphatases, are considered key drug targets in
human disease. Currently, indirect proxies are used to estimate activity perturbations of kinases and
phosphatases associated with disease development. Although useful, these proxies do not provide a direct
measure of enzymatic activity, leading to inaccurate estimates of kinase and phosphatase activity. As a result,
a clear understanding of the role of kinase and phosphatase activity perturbations during the development and
progression of human disease is lacking. Therefore, there is a critical need for the development and application
of chemical tools to directly quantify kinase and phosphatase activity in human disease states. In this proposal,
we will leverage a phosphorylation-sensitive amino acid to construct a panel of kinase and phosphatase
activity probes and apply this panel to develop longitudinal activity profiles of signaling changes during human
disease progression.
 The finely tuned activity of signaling proteins is essential for normal physiology. Indeed, perturbations
in the activity of signaling proteins are central to the development of human disease. Unfortunately, the field
still lacks a unified approach to modulating the activity of multiple signaling proteins simultaneously in living
systems in order to model human disease. To address this critical need, we will leverage split-protein
reassembly in order to define a lexicon of standardized parts for fine-tuning the activity of signaling nodes in
living cells. In the long-term, this set of mix-and-match parts will be utilized to model human disease states and
identify potential drug targets.
 Lastly, our laboratory will investigate the fundamental aspects of protein misfolding, which is now
recognized as a central pathological mechanism in numerous human disease states. The current lack of
approaches to assess protein misfolding and aggregation in living systems has created a critical need for the
development of novel methodologies to address this gap. By leveraging a novel, luminescence-based assay
for protein misfolding and aggregation our laboratory will assess the molecular determinants of protein
aggregation in living cells. In addition, we will utilize this approach to identify molecules capable interfering with
protein aggregation.

## Key facts

- **NIH application ID:** 9934242
- **Project number:** 5R35GM119751-06
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Cliff I Stains
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $343,392
- **Award type:** 5
- **Project period:** 2016-07-15 → 2022-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9934242

## Citation

> US National Institutes of Health, RePORTER application 9934242, Chemical Approaches for Interrogating Fundamental Biomedical Processes (5R35GM119751-06). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/9934242. Licensed CC0.

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