# Purchase of helium recovery system for NMR spectrometers

> **NIH NIH R01** · TEXAS A&M AGRILIFE RESEARCH · 2020 · $224,958

## Abstract

The RING ubiquitin E3 ligases are a superfamily of proteins critical to protein homeostasis and signaling in
eukaryotes. Dysfunctions in E3 ligases are implicated in innumerable human diseases. This proposal focuses
on the regulation of the ubiquitin E3 ligase Parkin. Parkin is central to the controlled destruction of damaged
mitochondria by autophagy (mitophagy). Controlled mitophagy is particularly essential to cardiac and
neuronal health. Uncontrolled mitophagy due to mutations in Parkin is clearly a driver of early onset
Parkinson's disease (eoPD). Parkin is now implicated in a number of other neurological diseases,
cardiomyopathy and in various cancers. The central goal here is to create an understanding the physical basis
for regulation of Parkin and how clinically observed mutations promote unregulated activity leading to
inadequately controlled mitophagy and other biological defects.
Though much is known about the biology and structural basis of Parkin function, very little is certain about
the physical basis for its regulation. Parkin activity is suppressed by its intra-molecular association with a
ubiquitin-like domain and is allosterically activated by the binding of phosphorylated ubiquitin (pUb).
Phosphorlyation of the Ubl domain also promotes activation. This complicated intersection of regulatory
mechanisms can only be understood by the rigorous dissection of the underlying thermodynamics. Without
this knowledge one cannot fully interpret the effects of mutations that lead to disease.
We shall take advantage of the broad foundation of knowledge of the biology of Parkin and structural basis of
its function to address the poorly understood thermodynamics of allosteric regulation of Parkin. The basis for
regulatory control of Parkin will be cast in a modern statistical thermodynamics description of the protein
ensemble. The influence of allosteric regulators and post-translational modifications will be examined by
comprehensive hydrogen exchange monitored by mass spectrometry and NMR spectroscopy; advanced NMR
relaxation techniques; single molecule fluorescence; calorimetry; enzymology; and mutagenesis.
A more rigorous and complete understanding of the regulation of Parkin will enable a robust interpretation
of pathological mutations. Not all pathological mutations can be simply explained as mutations that disrupt
the levels of protein or mutations that directly impact the catalytic site. Examples of common pathological
mutations will be examined to reveal the basis for their effects on Parkin's regulatory fidelity, with a longer-
range goal of determining how this impact might be mitigated by small molecule intervention.

## Key facts

- **NIH application ID:** 10135423
- **Project number:** 3R01GM129076-01A1S1
- **Recipient organization:** TEXAS A&M AGRILIFE RESEARCH
- **Principal Investigator:** A. JOSHUA WAND
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $224,958
- **Award type:** 3
- **Project period:** 2020-03-01 → 2024-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10135423

## Citation

> US National Institutes of Health, RePORTER application 10135423, Purchase of helium recovery system for NMR spectrometers (3R01GM129076-01A1S1). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10135423. Licensed CC0.

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