1H nuclear magnetic resonance assay of erythrocyte triosephosphate isomerase

• Published in: Analytical biochemistry Vol. 197; no. 1; pp. 178 - 181
• Main Authors: Hyslop, Serena J., Beal, Patricia, Kuchel, Philip W.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 15.08.1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Enzymes and enzyme inhibitors; Erythrocytes - enzymology; Fundamental and applied biological sciences. Psychology; Hemolysis; Heterozygote; Homozygote; Humans; Hydrogen; Magnetic Resonance Spectroscopy - methods; Triose-Phosphate Isomerase - blood; Triose-Phosphate Isomerase - deficiency; Triose-Phosphate Isomerase - genetics; Human; Proteins; Enzymatic activity; Enzyme; Triosephosphate isomerase; Red blood cell; NMR spectrometry
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(91)90375-4

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.