Orientation of cytochrome c oxidase molecules in the two populations of reconstituted vesicles resolved by column chromatography on DEAE-Sephacryl.
Vesicles reconstituted with bovine heart cytochrome c oxidase and dioleoylphosphatidylcholine can be resolved into two populations by column chromatography in DEAE-Sephacryl (Madden, T.D. and Cullis, P.R. (1984) J. Biol. Chem. 259, 7655-7658). These two fractions (I and II) were treated with two proteases. These are trypsin, which has been found to cleave subunit IV in the M domain of the cytochrome c oxidase molecule, and chymotrypsin, which has been found to cleave subunit III in the C domain. These studies show that fraction I vesicles contain cytochrome c oxidase orientation with the M domain outside, i.e., in the same topology as in submitochondrial particles, while fraction II vesicles contain enzyme molecules with their C domain outside, and thus in the same orientation as in mitochondria.
Zhang
Y Z
YZ
Capaldi
R A
RA
Cullis
P R
PR
Madden
T D
TD
D000818:Animals; D002417:Cattle; D002845:Chromatography; D002918:Chymotrypsin / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000032:analysis*; D008929:Mitochondria, Heart / Q000201:enzymology; D014357:Trypsin / Q000494:pharmacology
D003576:Electron Transport Complex IV; D002918:Chymotrypsin; D014357:Trypsin