Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.704621
Title: Studies on rabbit liver phosphoglucomutase
Author: Jamil, Haris
Awarding Body: University of London
Current Institution: Royal Holloway, University of London
Date of Award: 1983
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Abstract:
Phosphoglucomutases from different sources exhibit a variety of kinetic behaviour though it is probable they all have a common phosphoenzyme mechanism. The variations in behaviour arise partly from an intrinsic diphosphatase activity, which, even when present as a minor component, can affect the initial velocity patterns. Only isotopic induced-transport tests distinguish unequivocally between possible mechanisms. Rabbit liver phosphoglucomutase has a substantial amount of an isoenzyme, which appears to be associated with a high intrinsic diphosphatase activity. This activity may be involved in modulation of the glucose-1,6-diP level in the liver and hence in the regulation of carbohydrate metabolism. A procedure for the isolation of comparatively undegraded phosphoglucomutase (mol. wt. 67,600) from rabbit liver with high specific activity is described. The purified preparation showed a low diphosphatase activity. Kinetic studies, using a sensitive fluorimetric assay at ionic strength 0.05 mol 1-1 and Mg++ free = 2.0 mM, give a parallel line initial velocity pattern. This behaviour is consistent with the phosphoenzyme mechanism. The liver enzyme indicated a higher Km for glucose-1,6-diP than many other phosphoglucomutases. Induced transport tests using 14C- and 32P-labelled substrates showed that the enzyme can only have a phosphoenzyme mechanism. The isolation of 32P-labelled phosphoenzyme which rapidly exchanged more than 95% of its 32P-label with substrates confirmed that this phosphoenzyme is a true kinetic intermediate in the mutase reaction. Labelled phosphoenzyme contained 0.45-0.6 mol 32P mol-1 enzyme and its half-life was 47.5 h at 30 °C. Evidence is presented that the phosphate is bound to a serine residue of the enzyme. It is concluded that the low diphosphatase activity associated with the enzyme does not significantly modulate the level of glucose-1,6-diP in liver. Moreover, other factors must be responsible for the failure to isolate a completely phosphorylated phosphoenzyme. Although this phosphoglucomutase has a relatively high Km for glucose-1,6-diP, the known fluctuations in the level of glucose-1,6-diP in the liver are unlikely to affect phosphogluco-mutase activity.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.704621  DOI: Not available
Keywords: Biochemistry
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