Investigating ligand interactions and dynamics on the membrane proteome through membrane mimetic thermal proteome profiling (MM-TPP)
department or school: Medicine, Faculty of
department or school: Biochemistry and Molecular Biology, Department of
data provider: DSpace
degree campus: UBCV
creator: Jandu, Rupinder
issued: 2025-04-10T18:29:26Z
issued: 2025
related degree: Master of Science - MSc
degree grantor: University of British Columbia
description: Membrane proteins (MPs) are central to numerous cellular processes and represent key targets in drug discovery, yet their characterization can be challenging due to their hydrophobic nature and reliance on detergents for solubilization. Traditional workflows may destabilize MPs, hindering the study of their ligand interactions and downstream effects. In this thesis, we develop a novel membrane mimetic-based thermal proteome profiling (MM-TPP) workflow that aims to overcome these challenges by enabling the investigation of ligand-specific interactions on membrane proteomes in a detergent-free environment. Using peptidisc, we successfully capture integral membrane proteins (IMPs) into water-soluble libraries, facilitating downstream thermal proteome profiling. Using ATP and vanadate together (ATP-VO₄) as ligands, we validate the approach in both bacterial cells and mouse organ. Our results highlight ATP-VO₄-mediated stabilization of IMPs such as the bacterial ABC transporters MsbA and LolCD. As well as liver transporters like FATP2/SLC27A2 and BSEP/ABCB11. Beyond small-molecule ligands, MM-TPP highlights the effects of co-captured lipids on protein stability, underscoring the broader scope of this methodology. Additionally, MM-TPP has the potential to detect off-target stabilization effects of ATP derivatives, such as ADP, on non-ATP-binding proteins. showcasing its ability to uncover ligand metabolism and downstream interactions. This thesis establishes an introduction to MM-TPP as a basis for membrane proteome characterization.
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/90626
note: INVESTIGATING LIGAND INTERACTIONS AND DYNAMICS ON THE MEMBRANE PROTEOME THROUGH MEMBRANE MIMETIC THERMAL PROTEOME PROFILING (MM-TPP) by Rupinder Jandu B.Sc., McMaster University, 2022 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Biochemistry and Molecular Biology) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) April 2025 © Rupinder Jandu, 2025 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the thesis entitled: INVESTIGATING LIGAND INTERACTIONS AND DYNAMICS ON THE MEMBRANE PROTEOME THROUGH MEMBRANE MIMETIC THERMAL PROTEOME PROFILING (MM-TPP) submitted by Rupinder Jandu in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry and Molecular Biology Examining Committee: Dr. Frank Duong van Hoa, Professor, Biochemistry and Molecular Biology, UBC Supervisor Dr. Seth Parker, Professor, Biochemistry and Molecular Biology, UBC Supervisory Committee Member Dr. Thibault Mayor, Professor, Biochemistry and Molecular Biology, UBC Supervisory Committee Member Dr. Robert Molday, Professor, Biochemistry and Molecular Biology, UBC Additional Examiner iii Abstract Membrane proteins (MPs) are central to numerous cellular processes and represent key targets in drug discovery, yet their characterization can be challenging due to their hydrophobic nature and reliance on detergents for solubilization. Traditional workflows may destabilize MPs, hindering the study of their ligand interactions and downstream effects. In this thesis, we develop a novel membrane mimetic-based thermal proteome profiling (MM-TPP) workflow that aims to overcome these challenges by enabling the investigation of ligand-specific interactions on membrane proteomes in a detergent-free environment. Using peptidisc, we successfully capture integral membrane proteins (IMPs) into water-solu…
has type: Thesis/Dissertation
date issued: 2025-05
is shown at: 10.14288/1.0448349
language: eng
degree discipline: Biochemistry and Molecular Biology
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Duong, Franck
type: Text
identifier URI: http://hdl.handle.net/2429/90626