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Title: Some studies in purine metabolism and control
Author: Crozier, David Henry
ISNI:       0000 0001 3399 0467
Awarding Body: University of Warwick
Current Institution: University of Warwick
Date of Award: 1974
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The flux of metabolites within the cell is discussed. Two approaches were used in the study of this subject: 1. A preliminary investigation of the controlling factors of adenine metabolism in red blood cells was undertaken. 2. A study was made of adenylate cyclase in red blood cells and of the cyclic-AMP binding proteins of the bovine adrenal cortex. AMP deaminase was partially purified from human and sheep erythrocytes. This greatly facilitated the spectrophotometric assay of the enzyme’s activity. Some properties of this enzyme were studied. A method for examining the control of purine flux was developed by the use of radioisotopic tracers. The possibility of its use as a diagnostic assay for metabolic disorders is discussed. Adrenalin, which modifies the erythrocyte membrane, was tested for any effect it might have on the rate of transport of adenine into the red blood cell. The adrenal in or fluoride ion stimulated activities of adenylate cyclase were measured in turkey, rat and human red blood cells. The assay used was the saturation analysis procedure of Brown et al. (1971) using a cyclic-AMP binding protein from the bovine adrenal cortex. The cyclic-AMP binding moiety is the cyclic-AMP dependent protein kinase regulatory subunit. The crude preparation used in the assay for cyclic-AMP was fractionated by several methods and cyclic-AMP binding studies were carried out on the different fractions. Particular attention was paid to the interpretation of binding data. It was found that under different conditions certain preparations of the binding protein could produce either linear or curved binding plots ('bound' against 'bound/free’ ligand) The causes of this are discussed. The result of the binding study has mechanistic implications.
Supervisor: Not available Sponsor: Science Research Council
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: QD Chemistry