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An investigation into the expression and physiological function of mitochondrial uncoupling proteins
Carroll, Audrey
THESIS 7419 Our study provides evidence that rat thymus contains a functioning mitochondrial uncoupling protein (UCP 1). Using reverse transcriptase-polymerase chain reaction (RT-PCR), we showed that rat thymus contains UCP 1 mRNA. Antipeptide UCP 1 antibodies, that we show to be specific for UCP 1, detected UCP 1 protein in thymus and thymus cell (i.e. thymocyte) mitochondria. We also show that starvation (48-hour) induced no significant change in the abundance of UCP 1 in thymus and thymocyte mitochondria. Evidence for functional UCP 1 in rat thymus mitochondria was obtained using a radiolabeled binding assay in thymus and BAT mitochondria. UCP 1 in thymus mitochondria binds [3H] GDP with similar binding parameters, Bmax and KD values, as BAT mitochondria. In addition, a large component of the non-phosphorylating oxygen consumption by thymus mitochondria was inhibited by purine nucleotides and activated by nanomolar amounts of fatty-acids, akin to UCP 1 in BAT mitochondria. We also purified UCP 1 from thymus mitochondria by hydroxyapatite chromatography and identified the isolated protein by peptide mass mapping and tandem mass spectrometry using MALDI-TOF and LC-MS/MS, respectively. We conclude that rat thymus contains a functioning UCP 1 protein, which has the capacity to regulate metabolic flux and production and transport of reactive oxygen containing molecules in the thymus.
Keyword(s): Biochemistry, Ph.D.; Ph.D. Trinity College Dublin
Publication Date:
2004
Type: Doctoral thesis
Peer-Reviewed: Unknown
Language(s): English
Institution: Trinity College Dublin
Citation(s): Audrey Carroll, 'An investigation into the expression and physiological function of mitochondrial uncoupling proteins', [thesis], Trinity College (Dublin, Ireland). School of Biochemistry and Immunology, 2004, pp 267
Publisher(s): Trinity College (Dublin, Ireland). School of Biochemistry and Immunology
Supervisor(s): Porter, Richard
First Indexed: 2019-05-16 06:12:29 Last Updated: 2019-05-16 06:12:29