TY - JOUR
T1 - Cell-free analysis of targeting of cytochrome P450 to microsomal membranes
AU - Szczesna-Skorupa, Elzbieta
AU - Kemper, Byron
PY - 1991/1/1
Y1 - 1991/1/1
N2 - 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.
AB - 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.
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U2 - 10.1016/0076-6879(91)06077-G
DO - 10.1016/0076-6879(91)06077-G
M3 - Article
C2 - 1784247
AN - SCOPUS:0026345505
SN - 0076-6879
VL - 206
SP - 64
EP - 75
JO - Methods in enzymology
JF - Methods in enzymology
IS - C
ER -