Rat thiopurine methyltransferase assay procedure, developmental changes and strain variation

Randall C. Walker, Lee C. Woodson, Richard M Weinshilboum

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Abstract

Thiopurine methyltransferase (TPMT) is one of the enzymes involved in the catabolism of 6-mercaptopurine and azathioprine. An assay procedure was developed for rat TPMT, and the activity of the enzyme was measured in erythrocyte lysates and in homogenates of three solid tissues of Sprague-Dawley rats. Apparent Michaelis-Menten (Km) values for the two co-substrates of the reaction, 6-mercaptopurine (6-MP) and S-adenosyl-L-methionine (SAM), were 2.4, 1.8, 1.4 and 1.5 × 10-3M for 6-MP and 5.3, 3.3, 4.3 and 4.3 × 10-6M for SAM in erythrocyte, intestine, kidney, and spleen respectively. The pH optima were 6.7 in all four tissues. Sprague-Dawley rat kidney TPMT activity increased 28-fold and spleen activity increased 2-fold from birth to adulthood when expressed per gram of tissue. The increases were 13- and 2.2-fold, respectively, when expressed per milligram of protein. Enzyme activity was measured in tissues of adult male animals of nine inbred and two outbred rat strains. The rank order of tissue TPMT activity in all strains was: kidney > intestine > spleen > blood. There was a significant correlation between relative erythrocyte and relative kidney enzyme activities. Two-fold variations among strains in renal TPMT activity were found.

Original languageEnglish (US)
Pages (from-to)115-121
Number of pages7
JournalBiochemical Pharmacology
Volume30
Issue number2
DOIs
StatePublished - Jan 15 1981

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thiopurine methyltransferase
Rats
Assays
6-Mercaptopurine
Tissue
Kidney
S-Adenosylmethionine
Spleen
Erythrocytes
Enzyme activity
Enzymes
Intestines
Sprague Dawley Rats
Azathioprine
Animals
Blood
Parturition
Substrates

ASJC Scopus subject areas

  • Pharmacology

Cite this

Rat thiopurine methyltransferase assay procedure, developmental changes and strain variation. / Walker, Randall C.; Woodson, Lee C.; Weinshilboum, Richard M.

In: Biochemical Pharmacology, Vol. 30, No. 2, 15.01.1981, p. 115-121.

Research output: Contribution to journalArticle

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