The acylphosphate present in chromaffin granule membrane preparations is not associated with the proton-pump.
Chromaffin granule membranes were incubated in the presence of low ATP concentrations, at low temperature. A phosphorylated compound was rapidly formed which was stable in 10% trichloroacetic acid at 0 degree C. The lability of this compound in the presence of hydroxylamine or hot trichloroacetic acid indicated an acylphosphate, i.e., an ATPase phosphointermediate. Vanadate but not N-ethylmaleimide inhibited the formation of this derivative. Since the ATP-dependent generation of a transmembrane potential in chromaffin granule vesicles by the H+-pump was inhibited by N-ethylmaleimide but not by vanadate, the acylphosphate should not be associated with the H+-pump, i.e. ATPase I. We suggest that it is associated with ATPase II, an ATPase of unknown function present in chromaffin granule membrane preparations. This hypothesis is supported by the fact that ATPase II is vanadate sensitive and has a molecular mass of 140 kDa, properties similar to those of the phosphorylated intermediate.
D000255:Adenosine Triphosphate / Q000378:metabolism; D002837:Chromaffin Granules / Q000378:metabolism*; D002838:Chromaffin System / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D006898:Hydroxylamines / Q000494:pharmacology; D007425:Intracellular Membranes / Q000378:metabolism; D010766:Phosphorylation; D006180:Proton-Translocating ATPases / Q000378:metabolism*; D014639:Vanadium / Q000494:pharmacology
D006898:Hydroxylamines; D014639:Vanadium; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D005033:Ethylmaleimide