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A fluorimetric assay of the phosphoribosylation of 6-mercaptopurine in human blood cells.

P C Wong·Y Hey·J S Fowler
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Abstract

A new fluorimetric method to assay HGPRT (hypoxanthine-guanine phosphoribosyl transferase, EC 2.4.2.8) from human red cell lysates using 6MP (6-mercaptopurine) as substrate is described. This assay is compared with an existing spectrophotometric method using hypoxanthine as substrate. Precision, sensitivity limits and kinetic differences of these assays are reported.

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ErythrocytesHumansHydrogen-Ion ConcentrationHypoxanthine PhosphoribosyltransferaseKineticsMercaptopurineSpectrometry, FluorescenceJournal Article

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BibTeXRIS

P C Wong, Y Hey, J S Fowler. 1979-11-15. A fluorimetric assay of the phosphoribosylation of 6-mercaptopurine in human blood cells.. https://doi.org/10.1016/0009-8981(79)90138-4

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Erythrocytes↗
PubMed1979

Human hypoxanthine-guanine phosphoribosyltransferase. IMP-GMP exchange stoichiometry and steady state kinetics of the reaction.

Steady state kinetics of hypoxanthine guanine phosphoribosyltransferase-catalyzed reactions are studied. The results obtained suggest that IMP, GMP, P-Rib-PP, and pyrophosphate bind to the same enzyme form, while hypoxanthine and guanine bind to a different form. Guanine activates IMP-pyrophosphorolysis. During the reaction guanine and IMP are consumed with formation of GM

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