Studies on the relationship between the inner and outer membranes of rat liver mitochondria as determined by subfractionation with digitonin.
The present investigation has attempted to define in rat liver mitochondria the distribution of outer membrane proteins in relation to the inner membrane by fractionation with digitonin and phospholipase A2. Porin, the channel-forming protein in the outer membrane, was measured quantitatively by immunological methods. Neither monoamine oxidase nor porin could be released by phospholipase A2 treatment, but both were released by digitonin, at the same detergent concentration. Thus, the release of monoamine oxidase and porin requires the disruption of the cholesterol but not the phospholipid domains of the membrane and the two polypeptides exist in the same, or similar, membrane environment with regard to cholesterol. Changes in the energy state, or binding of brain hexokinase to rat liver mitochondria prior to fractionation with digitonin, did not alter the release patterns of porin and monoamine oxidase. The uptake of Ca2+, however, resulted in the concomitant release of the outer membrane markers together with the matrix marker, malate dehydrogenase. The present findings with liver differ from those obtained recently with brain mitochondria (L. Dorbani et al. (1987) Arch. Biochem. Biophys. 252, 188-196) in which two populations of porin were located in two different cholesterol domains. The significance of these differences in the location of porin in liver and brain mitochondria is discussed.
Jancsik
V
V
Institute of Enzymology, Hungarian Academy of Sciences.
Lindén
M
M
Dorbani
L
L
Rendon
A
A
Nelson
B D
BD
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D002458:Cell Fractionation; D002784:Cholesterol / Q000378:metabolism; D004072:Digitonin; D007425:Intracellular Membranes / Q000201:enzymology / Q000378:metabolism*; D008291:Malate Dehydrogenase / Q000378:metabolism; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000201:enzymology / Q000378:metabolism*; D018272:Porins; D051381:Rats; D011919:Rats, Inbred Strains
D001425:Bacterial Outer Membrane Proteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D018272:Porins; D002784:Cholesterol; D008291:Malate Dehydrogenase; D004072:Digitonin