Clinical Chemistry
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Clinical Chemistry 28: 2026-2032, 1982;
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Clinical Chemistry, Vol 28, 2026-2032, Copyright © 1982 by American Association for Clinical Chemistry

Gas-chromatographic assay for heme oxygenase activity

FW Sunderman Jr, JR Downs, MC Reid and LM Bibeau

We have developed an improved assay for microsomal heme oxygenase activity, based on the enzymic release of CO from the alpha-methene bridge of hemin and the quantitation of CO by gas chromatography. The within-run coefficient of variation (CV) of heme oxygenase assays in microsomes from rat tissues (liver, kidney) averaged 8%; the between- run CV averaged 15%. The detection limit for heme oxygenase activity was approximately 1 nmol/h per milligram of microsomal protein. Gas- chromatographic assays of heme oxygenase activities in rat tissues correlated well (r = 0.94) with results by a spectrophotometric assay based on bilirubin production. In untreated rats, heme oxygenase activity averaged 7 +/- 3 nmol/h per milligram of protein (n = 36) in kidney microsomes and 14 +/- 5 nmol/h per milligram of protein (n = 17) in liver microsomes. Heme oxygenase activity was increased 10-fold in kidney microsomes and threefold in liver microsomes from rats killed 17 h after subcutaneous injection of NiCl2 (0.5 mmol/kg body wt). These findings illustrate the efficacy of the gas-chromatographic assay for measuring xenobiotic effects on heme oxygenase activity.


The following articles in journals at HighWire Press have cited this article:


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 Am. J. Physiol. Heart Circ. Physiol.Home page
Y. Morimoto, W. Durante, D. G. Lancaster, J. Klattenhoff, and F. K. Tittel
Real-time measurements of endogenous CO production from vascular cells using an ultrasensitive laser sensor
Am J Physiol Heart Circ Physiol, January 1, 2001; 280(1): H483 - H488.
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Copyright © 1982 by the American Association for Clinical Chemistry.