# Elucidating the Molecular Basis of Cellular Metal Stress by using Mass Spectrometry-Based Proteomic Methods

> **NIH NIH R01** · DUKE UNIVERSITY · 2024 · $85,619

## Abstract

PROJECT SUMMARY/ABSTRACT
Requested here are funds to purchase FPLC instrumentation that will support NIH-funded research in an R01
grant (1R01GM145035-01A1) entitled, “Elucidating the Molecular Basis of Cellular Metal Stress by using
Mass Spectrometry-Based Proteomic Methods.” The instrument to be purchased is a Cytiva AKTA Pure
Fast Protein Liquid Chromatography System (FPLC). The requested funds will be used to purchase the
instrument and associated attachments necessary for its function (approximately $85,000). Access to this
instrument is critical to the success of research being performed. The on-going and proposed studies related to
the above grant build on preliminary and recently published results from this collaborative team showing the
utility of a pulse proteolysis mass spectrometry method developed in MPI Fitzgerald’s laboratory to identify
protein targets of Cu in E. coli, establishing these proteomic methodologies for the study of metal-protein
interactions. These preliminary and recently published results have identified protein targets that are now
being isolated and purified for rigorous mechanistic studies. The fplc instrumentation is critical for the rigor of
protein purification needed for functional and spectroscopic studies. The overall objective of the parent grant is
to identify proteins that are functionally affected when cells experience stress induced by exposure to excess
levels of Zn, Cu, and Ag, as a non-native surrogate of Cu(I). This objective is being met by using a powerful
combination of mass spectrometry-based proteomic methods to address four specific aims: 1) Determine
global profiles of protein stability changes as a function of cellular metal overload and metal deficiency across
bacterial, fungal, and human cancer cells; 2) Establish a mechanistic basis linking differential stability of Aim 1
proteins to function; 3) Identify the relative sensitivity of proteins across the proteome to misfolding induced by
Cu, Ag, and Zn binding; and 4) Establish a biophysical basis for understanding the relative sensitivity of
proteins to metal-induced misfolding. Two of the four above specific aims (i.e., Aims 2, and 4) require access to
the requested instrument.

## Key facts

- **NIH application ID:** 11099412
- **Project number:** 3R01GM145035-03S2
- **Recipient organization:** DUKE UNIVERSITY
- **Principal Investigator:** Michael C Fitzgerald
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $85,619
- **Award type:** 3
- **Project period:** 2022-04-05 → 2026-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/11099412

## Citation

> US National Institutes of Health, RePORTER application 11099412, Elucidating the Molecular Basis of Cellular Metal Stress by using Mass Spectrometry-Based Proteomic Methods (3R01GM145035-03S2). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/11099412. Licensed CC0.

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