The Ca2+-binding glycoprotein as the site of metabolic regulation of mitochondrial Ca2+ movements.

A change in the redox state of pyridine nucleotides such as that evoked by addition of oxaloacetate has been shown to promote Ca2+ efflux from Ca2+ pre-loaded respiring mitochondria. An affinity-chromatography-purified antibody preparation obtained against the mitochondrial Ca2+-binding glycoprotein inhibits the phenomenon. This finding suggests that the glycoprotein is involved also in the oxaloacetate-induced Ca2+ release. This conclusion is reinforced by the finding that Ca2+-binding glycoprotein shows four sites per molecule where the pyridine nucleotides may be bound. Binding of NAD+ occurs preferentially over the others and the binding shows positive cooperativity, indicating that the glycoprotein undergoes an allosteric change upon NAD+ binding. Interestingly, in addition, NAD+ lowers the affinity of the glycoprotein for Ca2+. The effect cannot be induced by NADH. Pyridine nucleotide phosphates, NADP+ and NADPH, are essentially not bound. The results are consistent with the view that the glycoprotein is the site of regulation of Ca2+ equilibration across the mitochondrial membrane and make it possible to conclude that the effector in the phenomenon is NAD+.