Critical Amino-terminal Segments in Insertion of Rat Liver Cytochrome P450 3A1 into the Endoplasmic Reticulum Membrane

Overview

Journal Experientia

Specialty Science

Date 1996 Sep 15

PMID 8841512

Citations 2

Authors

P J van den Broek

M Barroso

M C Lechner

Affiliations

Soon will be listed here.

Abstract

An in vitro transcription-translation assay was used to study the membrane topology of rat liver cytochrome P450 3A1. N-terminus deletion mutants were constructed to assess the importance of N-terminal regions in the stable incorporation of the protein into the microsomal membranes. Wild-type nascent cytochrome P450 bound to microsomes as an integral membrane protein through its hydrophobic N-terminal segments, uncleaved by signal peptidase. Deletion of the most N-terminal hydrophobic segment (positions 7-26) had a dramatic effect on endoplasmic reticulum membrane integration. Confirming the essential role of this stretch in P450 3A1 membrane targeting, proteolysis-resistant membrane-associated peptides were observed in all the in vitro translated mutants containing that segment. It is concluded that the membrane topogenesis of P450 3A1 is determined mainly by the amino-terminal hydrophobic segment.

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