A study of ornithine aminotranferase and intracellular orinthine metabolism

Leah, J.M., 1988. A study of ornithine aminotranferase and intracellular orinthine metabolism. PhD, Nottingham Trent University.

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Abstract

The mitochondrial matrix enzyme, ornithine aminotransferase, has been purified from rat and human sources. The enzyme was found to undergo self-association upon concentration, in agreement with the findings of previous workers. Native molecular weight studies in the absence and presence of detergent suggest that the hydrophobicity and aggregation of ornithine aminotransferase may spare a high concentration intramitochondrial ornithine pool from metabolism by this enzyme. Characterization of rat and human ornithine aminotransferase has shown them to be very similar. Polyclonal antibodies have been raised to the enzyme from both sources and immunodiffusion analysis indicates a close structural similarity. This similarity has been exploited to develop an immunoadsorbent capable of purifying human ornithine aminotransferase using affinity purified rabbit antibodies. This technique has been successfully applied to human liver, rat kidney and rat blood platelet extracts. Results have shown that blood platelets may be a suitable tissue source of ornithine aminotransferase for future investigation of patients with Gyrate Atrophy, an inherited deficiency of this enzyme. A competitive enzyme-linked immunosorbent assay has been developed to detect low levels of ornithine aminotransferase and both rat and human tissues have been assayed. Estimations were in agreement with activity measurements and the monospecificity of the immunoassay has been checked by Western blotting.

Two analytical systems have been assessed for quantitative amino acid analysis of physiological fluids from patients with inborn errors of metabolism, in particular, samples from Gyrate Atrophy patients. Attempts have been made to investigate intracellular amino acid levels and the phenomenon of membrane-associated amino acids, in rat liver extracts. Ornithine was found to be undetectable in mitochondria. This could indicate very low levels in the mitochondrial matrix, or might be an artifact due to rapid diffusion out of organelles.

Item Type:	Thesis
Creators:	Leah, J.M.
Date:	1988
ISBN:	9781369324334
Identifiers:	Number Type PQ10290184 Other
Rights:	This copy of the thesis has been supplied on condition that anyone who consults it is understood to recognise that its copyright rests with its author and that no quotation from the thesis and no information derived from it may be published without the author's prior written consent
Divisions:	Schools > School of Science and Technology
Record created by:	Linda Sullivan
Date Added:	11 Nov 2020 15:33
Last Modified:	11 Oct 2023 10:22
URI:	https://irep.ntu.ac.uk/id/eprint/41622

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