# Engineered Nanodiscs for Structural Mass Spectrometry

> **NIH NIH R01** · UNIVERSITY OF MICHIGAN AT ANN ARBOR · 2022 · $341,184

## Abstract

Project Summary
 The overall objective of this project is to develop new microfluidic devices capable of
producing lipid nanodiscs (NDs) having detailed and tunable compositions, and utilize these
NDs as vehicles for improved structural mass spectrometry (SMS) and proteomics assays of
membrane proteins (MPs). Despite positions of prominence in both biochemistry and the
pharmaceutical sciences, our understanding of MPs lags significantly behind our knowledge of
soluble proteins and their functional complexes. This has led to a critical imbalance, where
MPs, which account for greater than 60% of drug targets, account for ~3% of the structural
entries in the protein data bank (PDB). As a result, MP-targeted drug discovery is slowed, and
treatment strategies go undiscovered.
 To bridge this gap, we propose the development of monolithic microfluidic tools capable
of producing NDs over a wide range of lipid compositions, having narrow size distributions. We
will then immediately deploy these tailored NDs to study the structure and biophysics of
cytochrome P450 (CYP), a monotopic membrane protein found in all kingdoms of life, and
responsible for the metabolism of most small molecule drugs in humans. The means by which
CYPs are able to metabolize such a wide range of xenobiotics, and the role that cellular
membranes play in the apparent structural plasticity of CYPs, remains unknown. We will
develop ion mobility-mass spectrometry (IM-MS) and collision induced unfolding (CIU) methods
that, in our preliminary data, have been able to detect the first evidence for structural shifts in
CYP as a function of its local lipid environment. Our efforts will further extend to build NDs into
robust extraction devices for improved coverage of the membrane proteome. To complement
our IM-MS workflows, we will also deploy and optimize chemical cross-linking (CXL)
approaches for use with MPs housed within NDs.
 The tools discussed above will then be applied to the study of the MP complexes within
the mitochondrial membrane. Specifically, we will target the protein assemblies associated with
the electron transport chain (ETC), as well as the vast array of protein-protein interactions
(PPIs) that have either been observed or predicted for CYP. Our work will seek to provide new
structural information on complexes that have been studied extensively in the past (e.g. ATP
Synthases), as well as seek to discover new mitochondrial MP complexes, with potentially
broad implications for cellular function.

## Key facts

- **NIH application ID:** 10460573
- **Project number:** 5R01GM138620-03
- **Recipient organization:** UNIVERSITY OF MICHIGAN AT ANN ARBOR
- **Principal Investigator:** Philip C Andrews
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $341,184
- **Award type:** 5
- **Project period:** 2020-09-22 → 2024-07-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10460573

## Citation

> US National Institutes of Health, RePORTER application 10460573, Engineered Nanodiscs for Structural Mass Spectrometry (5R01GM138620-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10460573. Licensed CC0.

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