An amphitropic cAMP-binding protein in yeast mitochondria. 3. Membrane release requires both Ca2(+)-dependent phosphorylation of the cAMP-binding protein and a phospholipid-activated mitochondrial phospholipase.
The amphitropic cAMP-binding protein in mitochondria of the yeast Saccharomyces cerevisiae is released from the inner membrane into the intermembrane space by the degradation of its lipid membrane anchor consisting of or containing phosphatidylinositol. The releasing reaction depends on the presence of an N-ethylmaleimide-sensitive protein (releasing factor) in the intermembrane space and is controlled by Ca2+ and phospholipid (or lipid derivatives). Here we demonstrate that these two effector molecules act on different activation steps within a complex releasing pathway involving both the cAMP receptor and the releasing factor: Ca2(+)-dependent phosphorylation of the receptor protein seems to be prerequisite for its subsequent lipolytic liberation from the inner membrane. In the presence of phospholipid (or lipid derivatives) the previously soluble releasing factor, which may be identical with a soluble diacylglycerol-binding protein in the mitochondrial intermembrane space, associates with the inner membrane. This change in the intramitochondrial location of the releasing factor, which thus exhibits amphitropic behavior itself, may be required for (direct or indirect) activation of the mitochondrial phospholipase which then releases the cAMP receptor from the inner membrane in a form liable to dissociation from the C subunit by cAMP.
Müller
G
G
Institut für Pathologie und Rechtsmedizin, Universität Ulm, München, Federal Republic of Germany.
Bandlow
W
W
D002118:Calcium / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002373:Cyclic AMP Receptor Protein; D004789:Enzyme Activation; D007425:Intracellular Membranes / Q000378:metabolism; D008928:Mitochondria / Q000201:enzymology / Q000378:metabolism*; D010740:Phospholipases / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism / Q000494:pharmacology; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000502:physiology; D011953:Receptors, Cyclic AMP / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000648:ultrastructure*; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism
D002352:Carrier Proteins; D002373:Cyclic AMP Receptor Protein; D010743:Phospholipids; D011953:Receptors, Cyclic AMP; D011494:Protein Kinases; D010740:Phospholipases; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium