Thesis

Establishing a quantitative proteomic platform to profile mitochondrial-based diseases

Creator
Rights statement
Awarding institution
  • University of Strathclyde
Date of award
  • 2026
Thesis identifier
  • T17995
Person Identifier (Local)
  • 202475036
Qualification Level
Qualification Name
Department, School or Faculty
Abstract
  • Mitochondrial dysfunction, associated with compromised protein function within the respiratory chain, is frequently observed in ageing and can result in the onset of various neurodegenerative diseases. Deficiency of carnitine, a small molecule involved in fatty acid transport across the mitochondrial membranes, has recently been identified in frail persons. The carnitine shuttle system is composed of three proteins: carnitine palmitoyltransferase I (CPT1), carnitine palmitoyltransferase II (CPT2) and carnitine-acylcarnitine translocase (CACT). This project aims to apply an absolute quantification method to CPT2 whose role is to convert acylcarnitines present within the mitochondrial matrix to acyl-CoA that can be used for beta oxidation. Establishing a methodology to profile the abundance of these proteins in frail individuals may help the answer why the levels of carnitine decrease in frailty. In this project, workflow for peptide mapping was established using bovine serum albumin. This was then applied to full-length carnitine palmitoyltransferase II, which gave no observable results, presumably due to the hydrophobic nature of the membrane protein. Thus, truncated carnitine palmitoyltransferase II was profiled, which aided selection of a suitable peptide for quantification. In addition, hydrogen isotope exchange reactions to synthesise heavy isotope labelled amino acids that are compatible for incorporation by solid phase peptide synthesis were explored. Deuterated alanine was obtained in good yield and incorporated into a carnitine palmitoyltransferase II peptide, however, inexhaustive deuterium incorporation as well as overlap with the native peptide limit the applicability of the deuterated amino acid building block in proteomics studies. Absolute quantification of truncated carnitine palmitoyltransferase II was finally carried out using the absolute quantification (AQUA) method employing a heavy leucine labelled peptide.
Advisor / supervisor
  • Burley, Glenn A.
Resource Type
DOI
Embargo Note
  • This thesis is restricted to Strathclyde users only until 6th May 2031.

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