Abstract
Publisher Summary This chapter discusses the P450 phosphorylation in isolated hepatocytes and in vivo. Many P450 enzymes are regulated at the messenger RNA (mRNA) level, in response to changes in circulating levels of drugs and hormones. Studies in vitro have demonstrated that a number of hepatic P450s can be phosphorylated by purified protein kinases. Support for the occurrence of P450 phosphorylation under physiological conditions is provided by studies of P450 phosphorylation in isolated rat hepatocytes3–5 and in vivo. Several reports point to the potential importance of these phosphorylation reactions for the control of P450 activity, and perhaps also the regulation of P450 degradation. This chapter describes both in vitro and in vivo experimental protocols that have been developed for studies of P450 phosphorylation. Hepatocytes are isolated from untreated rats or from rats induced with drugs such as phenobarbital by perfusion of liver tissue with buffers containing collagenase using the nonrecirculating system described by Seglen.
Published Version
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