An h.p.l.c. method is described for the assay of protoporphyrinogen oxidase activity in rat liver. A relatively pure protoporphyrinogen IX substrate was obtained by selectively removing any protoporphyrin IX unreduced by sodium amalgam on a small disposable cartridge packed with a strong anion-exchanger. The protoporphyrin IX formed was extracted with dimethyl sulphoxide/methanol (3:7, v/v) containing mesoporphyrin as the internal standard for separation and quantification by reversed-phase chromatography. The Km for protoporphyrinogen was 9.5 +/- 1.6 microM, and the enzyme activities were 0.59 +/- 0.11 nmol of protoporphyrin IX produced/min per mg of mitochondrial protein and 33.5 +/- 2.7 nmol protoporphyrin IX produced/min per g of liver tissue homogenate. The method is applicable to the determination of enzyme activity in small amounts of human liver biopsy.
Research Article| May 01 1987
An h.p.l.c. assay for protoporphyrinogen oxidase activity in rat liver
C K Lim;
Biochem J (1987) 243 (3): 863–866.
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F Li, C K Lim, T J Peters; An h.p.l.c. assay for protoporphyrinogen oxidase activity in rat liver. Biochem J 1 May 1987; 243 (3): 863–866. doi: https://doi.org/10.1042/bj2430863
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