On the membrane topology of vertebrate cytochrome P-450 proteins.

Hydropathy profiles of 34 aligned cytochrome P-450 sequences were compared to identify potential transmembrane segments. Eleven regions with the potential to cross a membrane in at least some P-450 sequences were detected. The known sidedness of several residues and peptides was used to eliminate some of these regions from consideration. Further arguments based on the location and orientation of the heme relative to the membrane excluded others. This process of elimination was continued until only two regions remained. These two segments, present in the first 66 amino acids of the P-450 NH2 termini, are proposed as the only transmembrane peptides of vertebrate microsomal P-450s. Mitochondrial P-450s may have a different membrane association. The three-dimensional structure of cytochrome P-450cam was examined for the location of conserved charged residues. These residues occurred mainly on the opposite surface from the substrate-binding site and along the edges of the flat triangular P-450cam. A model is proposed for vertebrate microsomal P-450s that is similar to P-450cam. The substrate-binding site faces the membrane, the heme is parallel to the membrane surface, and two NH2-terminal transmembrane segments anchor the protein to the bilayer.