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Title: Expression and characterisation of the uncoupling proteins to yeast
Author: Harper, J. A.
Awarding Body: University of Cambridge
Current Institution: University of Cambridge
Date of Award: 2002
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Much of the evidence supporting a mitochondrial uncoupling function for the close homologues of UCP1 (e.g. UCP2 and UCP3) comes from experiments in which the levels of protein expression have been genetically manipulated. Evidence is presented, from yeast genetically modified to express UCP1 and UCP3, that these experimental manipulations can lead to an uncoupling of oxidative phosphorylation that does not necessarily represent a native activity of the protein. UCP1, whose native uncoupling activity can be fully inhibited by GDP, was used to demonstrate that artefactual uncoupling occurs at higher levels of protein expression. Similarly uncoupling was observed when UCP3 was expressed at high levels. UCP3 was expressed in yeast mitochondria at levels similar to, 2-fold and 7-fold greater than those found in mouse skeletal muscle (135 ± 45 ng/mg mitochondrial protein). No increase in uncoupling was seen than can be observed in fully inhibited UCP1. This uncoupling was not stimulated by the addition of palmitate nor was it inhibited by GDP. Therefore it is concluded that all uncoupling reported to be due to the expression of UCP3 is artefactual and does not represent a native activity of the protein.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available