Abstract
Procedures to remove noncytochrome haem or haemoglobin from rat liver microsomal preparations are described. Over 80% of the radioactive label due to exogenous [ 14C]haemoglobin could be removed by washing the microsomal suspension. In contrast, only about 50% of noncytochrome [ 14C]haem was removed by washing microsomes with either buffer or albumin solutions at 0 to 4°C. The most effective procedure to remove noncytochrome haem was to incubate the microsomal suspension for 5 to 6 min at 37°C in the presence of human serum albumin and dithioerythritol. Under these conditions, over 90% of the radioactive label was removed. This procedure brought about no reduction in the concentration of microsomal cytochrome b 5 or P-450, or the integrity of the mixed function oxidases, as judged by the hydroxylation of aniline or the N-demethylation of aminopyrene. Under similar conditions human apohemopexin was no more effective than albumin in bringing about the removal of haem from the microsomal suspension.
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