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1H nuclear magnetic resonance assay of erythrocyte triosephosphate isomerase

✍ Scribed by Serena J. Hyslop; Patricia Beal; Philip W. Kuchel

Publisher
Elsevier Science
Year
1991
Tongue
English
Weight
423 KB
Volume
197
Category
Article
ISSN
0003-2697

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✦ Synopsis

A direct method for measuring the activity of erythrocyte triosephosphate isomerase using 1H NMR spectroscopy was developed. NMR spectroscopy allows the simultaneous monitoring of the substrate and the product of the reaction by virtue of the differences in the NMR spectrum of each chemical species. The assay conditions were based on a modification of a conventional spectrophotometric method. The enzymatic activity measured using NMR gave results comparable to those obtained in a standard assay. The results were used in the kinetic characterization of triosephosphate isomerase in hemolysates from subjects with homozygous or heterozygous deficiency of the enzyme. In general, NMR spectroscopy has the potential for wide application in the rapid development of new enzyme assays.

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