The role of phospholipids in the modulation of enzyme activities in the chromaffin granule membrane.
(1) 93% of protein of chromaffin granule membranes can be solubilized by 1.3% (w/v) sodium cholate. The solubilized material can be substantially delipidated by ammonium sulphate precipitation. After three such cycles less than 2% of the endogenous phospholipids remain. (2) The chromaffin granule membrane Mg2+-ATPase depends on the presence of phospholipids for retention of its full activity. Soybean and extracted chromaffin granule phospholipids fully reactivate the delipidated enzyme provided only one delipidation step is used. (3) Successive ammonium sulphate precipitation steps result in a delipidated, and deactivated ATPase preparation which can be only partially reactivated on re-addition of phospholipids. (4) The phospholipid specificity for reactivation of the Mg2+-ATPase is broad. Although acidic phospholipids allow higher activities than neutral phospholipids, the main requirement appears to be the hydrophobic environment provided by the phospholipid hydrocarbon chains. (5) Correlations between changes in slope in the Arrhenius plot of the Mg2+-ATPase, and phase transitions in the phospholipid used for reactivation suggest that the 'fluidity' of the hydrocarbon chains modulates the activity of the enzyme.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000311:Adrenal Glands / Q000201:enzymology; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D002417:Cattle; D002793:Cholic Acids / Q000494:pharmacology; D002837:Chromaffin Granules / Q000201:enzymology*; D002838:Chromaffin System / Q000201:enzymology*; D004789:Enzyme Activation; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008563:Membrane Lipids / Q000502:physiology; D008565:Membrane Proteins / Q000502:physiology; D010743:Phospholipids / Q000494:pharmacology*; D013696:Temperature
D002793:Cholic Acids; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase