The reconstituted ADP/ATP carrier from mitochondria is both inhibited and activated by anions.
Anions were found to have a number of different effects on the reconstituted ADP/ATP carrier from mitochondria. (1) Binding of adenine nucleotides to the active site of the translocator is competitively inhibited by various anions. These anions can be arranged in a sequence of increasing competitive effect due to their order in a lyotropic series, and also due to increasing charge. (2) Apart from this competition effect, the presence of a sufficiently high concentration of anions turned out to be absolutely essential for functional ADP/ATP exchange in the reconstituted system. The activating anions too can be arranged in sequence, similar to that of the competition effect. The adenine nucleotide transport shows sigmoidal dependence on the stimulating anions with a Hill coefficient of n = 2. Addition of anions does not change the basic amount of functionally active translocator molecules. (3) The different effects of anions, i.e., inhibition and activation, were shown to take place at different sites and to be due to different mechanisms. Anions compete with substrates both at the outer (cytosolic) and at the inner (matrix) active site, whereas anion activation is observed solely by interaction with the cytosolic side of the translocator protein. (4) Activation of the reconstituted ADP/ATP exchange by anions could be discriminated from an activating influence of anionic phospholipids in the surroundings of the carrier protein.
D000818:Animals; D000838:Anions; D002412:Cations; D002417:Cattle; D007700:Kinetics; D008081:Liposomes; D008433:Mathematics; D008929:Mitochondria, Heart / Q000201:enzymology*; D000226:Mitochondrial ADP, ATP Translocases / Q000378:metabolism*; D008954:Models, Biological; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010743:Phospholipids / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013367:Submitochondrial Particles / Q000201:enzymology
D000838:Anions; D002412:Cations; D008081:Liposomes; D010743:Phospholipids; D000226:Mitochondrial ADP, ATP Translocases; D009713:Nucleotidyltransferases