Date of Award

1987

Degree Type

Dissertation

Degree Name

Ph.D.

Department

Chemistry and Biochemistry

Keywords

Chemistry, Biochemistry.

Rights

CC BY-NC-ND 4.0

Abstract

Use of bilirubin uridine 5$\sp\prime$-diphosphate glucuronyltransferase, for the analysis of bilirubin was studied. Microsomal suspensions, as well as the solubilized enzyme, demonstrated capacities for a kinetic assay of albumin: bilirubin (1:2) solutions and solutions devoid of albumin. The system was enzymatically coupled for continuous monitoring. Comparison with the Jendrassik-Grof method showed a very good agreement. Analysis of serum/plasma samples was met with interference from albumin. Albumin appears to inhibit the enzyme by sequestering free bilirubin, and possibly by direct protein-protein interaction with the enzyme. The low specificity of pyruvate kinase, for a phosphate acceptor, made it possible to link production of uridine 5$\sp\prime$-diphosphate to H$\sb2$O$\sb2$ production and bilirubin oxidation by peroxidase. The activity of bilirubin uridine 5$\sp\prime$-diphosphate glucuronyltransferase could be monitored continuously at 460 nm. Production of uridine 5$\sp\prime$-diphosphate is a feature of reactions catalyzed by uridine 5$\sp\prime$-diphosphate glucuronyltransferases. Therefore, the coupling system has the potential to allow assay of a number of UDPGT isoenzymes, as well as their substrates (which may be endogenous metabolites or xenobiotics). The pyruvate kinase can also be used for the analysis of adenosine 5$\sp\prime$-diphosphate, uridine 5$\sp\prime$-diphosphate and glucose.Dept. of Chemistry and Biochemistry. Paper copy at Leddy Library: Theses & Major Papers - Basement, West Bldg. / Call Number: Thesis1987 .S455. Source: Dissertation Abstracts International, Volume: 48-10, Section: B, page: 2963. Thesis (Ph.D.)--University of Windsor (Canada), 1987.

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