Microassay of heme oxygenase by high-performance liquid chromatography: application to assay of needle biopsies of human liver.
We developed a microassay for heme oxygenase, in which bilirubin (BR) production was measured by HPLC, and compared it to previously reported spectrophotometric methods. The microassay required as little as 5 mg wet human, rat, or chick embryo liver. Using the HPLC assay, values for heme oxygenase activity in extracts (10,000 g supernatant) of normal human liver obtained by needle biopsies were 44 +/- 7 (pmol BR.min-1.mg protein-1). Spectrophotometric assays of homogenates of human liver resulted in low values for heme oxygenase, due to unknown sources of interference. Comparative values of microsomal heme oxygenase activity were 294 +/- 25, 95 +/- 3, and 87 +/- 9 pmol BR.min-1.mg protein-1 for chick, rat, and human livers, respectively.
Lincoln
B C
BC
Department of Medicine, Emory University, Atlanta, Georgia 30322.
Mayer
A
A
Bonkovsky
H L
HL
D000818:Animals; D001663:Bilirubin / Q000096:biosynthesis; D001707:Biopsy, Needle; D002642:Chick Embryo; D002851:Chromatography, High Pressure Liquid; D006419:Heme Oxygenase (Decyclizing) / Q000032:analysis*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008832:Microchemistry; D008862:Microsomes, Liver / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000032:analysis*; D051381:Rats; D013045:Species Specificity; D013048:Specimen Handling
D006899:Mixed Function Oxygenases; D006419:Heme Oxygenase (Decyclizing); D001663:Bilirubin