# Understanding organization of membrane proteins and lipids through lipid vesicle native mass spectrometry

> **NIH NIH R01** · YALE UNIVERSITY · 2024 · $351,750

## Abstract

Abstract
In the crowded milieu of the membrane, membrane proteins, with other soluble and membrane-associated
proteins, and lipids form a large number of dynamic and transient protein complexes that in turn govern
cellular physiology. There is also mounting evidence that both independent membrane protein-lipid
interactions, as well as bulk biophysical properties of the host membrane often regulate these assemblies.
Hence, to understand how associations between specific membrane proteins help a cell responds to an
external stimulus, we need to study the oligomeric assemblies of the respective proteins directly from the
lipid bilayer environment. This brings us to the primary challenge of studying membrane protein-lipid
interactions. The existing tools to study such interactions lack this critical ability to perform molecular
analysis directly from the bilayer environment. Addressing this challenge, the overarching goal of this
project is to develop a novel experimental platform that enables analysis of MP complexes directly from in
vitro lipid bilayers, which can be customized to a target cellular membrane. To this end, we will combine
lipid vesicle technologies with native mass spectrometry (nativeMS). In Aim 1, taking a set of ten different
standard oligomeric membrane proteins we will develop an experimental method that enables us to
determine their oligomeric states directly from a range of lipid vesicles mimicking different physiological
membranes. We will validate and benchmark our results against the known oligomeric masses of each of
these proteins. This will establish the applicability of our platform to detect a wide range of membrane
proteins from a variety of lipid bilayer environments. In Aim 2, we will develop an experimental strategy that
enables us to directly determine the specifically bound lipid binds and where do they bind. To this end, in
collaboration with Thermo Fisher Scientific, we will combine ECD fragmentation with lipid vesicle nativeMS
platform. Together, upon successful completion, these two Aims will provide an arsenal of new
technologies to study the oligomeric organization of membrane proteins and lipids directly from a
physiologically relevant lipid bilayer. In Aim 3, we will apply this to a complex biological system to address
an outstanding question in neurobiology; how neurotransmitter filled synaptic vesicles attain their ultrafast
speed of fusion. To this end, we will specifically target the role of synaptophysin, a synaptic vesicle
membrane protein which has been linked to various neurological disorders. The experiment proposed can
bring out critical mechanist and structural insight to understand neuronal signal transduction and related
disease-specific impairments. In the long run, impairment of associations between membrane proteins has
been linked to several pathophysiological conditions ranging from neurodegeneration to cancer. We are
confident that the proposed platform will have a transformative role i...

## Key facts

- **NIH application ID:** 10834061
- **Project number:** 5R01GM141192-04
- **Recipient organization:** YALE UNIVERSITY
- **Principal Investigator:** Kallol Gupta
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $351,750
- **Award type:** 5
- **Project period:** 2021-05-01 → 2026-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10834061

## Citation

> US National Institutes of Health, RePORTER application 10834061, Understanding organization of membrane proteins and lipids through lipid vesicle native mass spectrometry (5R01GM141192-04). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10834061. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*