# Mechanism of protein aggregate recognition and disassembly by molecular chaperones

> **NIH NIH R01** · TEXAS A&M AGRILIFE RESEARCH · 2022 · $310,463

## Abstract

DESCRIPTION: The misfolding and aggregation of essential cellular proteins is a fundamental problem for all
living organisms. Aggregation of even non-essential proteins can lead to debilitating diseases like type II
diabetes, Alzheimer's, Huntington's and Parkinson's diseases. Importantly, protein folding and aggregation are
heavily influenced by the cellular protein quality control machinery, involving networks of molecular
chaperones. Precisely how different chaperone systems cooperate to dismantle and reactivate aggregated
proteins, and how molecular chaperone action affects disease progression, is not well understood. This
proposal will address three fundamental questions that impact this problem: First, what is the most accurate
physical description of protein aggregate disassembly by molecular chaperones? Second, in what way do the
structural properties of an aggregate nanoparticle impact how an they are taken apart? Third, how does the
critical small heat shock (sHsp) class of molecular chaperones enhance protein aggregate disassembly?
Addressing these questions in a detailed and quantitative manner is exceedingly difficult using standard
approaches, because the complex and heterogeneous nature of protein aggregates can obscure key
intermediates and transitions. Single particle analysis, in particular a fluorescence technique known as Burst
Analysis Spectroscopy (BAS), is ideally suited to overcome this problem. BAS can quantify complex
nanoparticle distributions in free solution, allowing for the detection of dynamically populated intermediates and
sub-populations. This project will employ BAS to study the disassembly of protein aggregates by two model
disaggregase systems, one from bacteria and one from yeast, at a level of detail unreachable by other
approaches. The overall goal is to develop a mechanistic understanding of how different molecular
chaperone networks recognize and dismantle protein aggregates that possess distinct physical
properties. In service of this goal, this project will extend the capabilities of BAS to incorporate multi-color and
Förster resonance energy transfer measurements. This project will also develop a set of new approaches that
are complementary to BAS and permit more detailed analysis of the hydrodynamic and structural properties of
aggregate nanoparticles by using (1) horizontal light sheet excitation and particle tracking in microfluidic flow
and (2) Tip-Enhanced Raman spectroscopy (TERS). It is anticipated that the combination of these techniques
will provide uniquely powerful approach to understanding protein disaggregation. Additionally, because the
core components of the chaperone networks examined in this work are conserved, it is further expected that
the discoveries made in these studies will contribute to a fundamentally better understanding of how molecular
chaperones recognize and process protein aggregates in human cells impacted by protein misfolding
diseases.

## Key facts

- **NIH application ID:** 10476451
- **Project number:** 5R01GM134063-04
- **Recipient organization:** TEXAS A&M AGRILIFE RESEARCH
- **Principal Investigator:** HAYS S RYE
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $310,463
- **Award type:** 5
- **Project period:** 2019-09-20 → 2025-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10476451

## Citation

> US National Institutes of Health, RePORTER application 10476451, Mechanism of protein aggregate recognition and disassembly by molecular chaperones (5R01GM134063-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10476451. Licensed CC0.

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