Cell-free analysis of targeting of cytochrome P450 to microsomal membranes

Elzbieta Szczesna-Skorupa, Byron Kemper

Research output: Contribution to journalArticlepeer-review


Analysis of P450 biosynthesis in cell-free systems has been useful in studying the mechanism of the targeting of P450 to the membrane (Table I). These techniques have allowed the demonstration that the N-terminal region of P450 is an uncleaved signal sequence for SRP-dependent membrane insertion. Lack of protection from protease indicated that most of the P450 is on the cytoplasmic side of the membrane. Substitution of basic amino acids at the P450 N terminus or replacing the N terminus of P450 with a secretory signal sequence results in partial or complete translocation across the membrane. This result indicates that at most one and probably none of the internal hydrophobic regions function as stop-transfer signals, which would be present if the protein spanned the membrane more than once.

Original languageEnglish (US)
Pages (from-to)64-75
Number of pages12
JournalMethods in enzymology
Issue numberC
StatePublished - Jan 1 1991
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


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