Separation of cytochrome P-450 containing vesicles from the midgut microsomal fraction of Manduca sexta.
1. Microsomes prepared from midgut tissue of Manduca sexta by differential centrifugation are a heterogeneous population of vesicles. 2. Upon centrifugation of microsomes in a sucrose gradient of 30%, 15% and 10%, specific activity of cytochrome P-450 catalyzed O-demethylation of p-nitroanisole (a marker enzyme for endoplasmic reticulum) was increased 2.5 to 3.5-fold in the 15% fraction. 3. Specific activities of alkaline phosphatase and leucine aminopeptidase (marker enzymes for brush border membranes) were increased 3.1 to 5.7-fold in the pellet. 4. Differential centrifugation is a useful step in the purification of cytochrome P-450 enzymes.
Purdue University, Department of Entomology, West Lafayette, IN 47907-1158.
J J
JJ
Neal
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Reuveni
D000818:Animals; D002499:Centrifugation, Density Gradient; D003577:Cytochrome P-450 Enzyme System / Q000302:isolation & purification*; D004064:Digestive System / Q000201:enzymology / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D008861:Microsomes / Q000201:enzymology; D008871:Microvilli / Q000201:enzymology; D009036:Moths / Q000201:enzymology*
D003577:Cytochrome P-450 Enzyme System