Phospholipase A(2) digestion of cardiolipin bound to bovine cytochrome c oxidase alters both activity and quaternary structure.
Phospholipase A(2) from Crotalus atrox hydrolyzes all of the phospholipids that are associated with purified, detergent-solubilized cytochrome c oxidase; less than 0.05 mol cardiolipin (CL)(1) remains bound per mol enzyme. Coincident with the hydrolysis of cardiolipin is a reversible decrease of 45-50% in the electron transport activity of the dodecylmaltoside-solubilized enzyme. Full activity is recoverable (90-98%) by addition of exogenous cardiolipin, but not by either phosphatidylcholine or phosphatidylethanolamine. Unexpectedly, cleavage of cardiolipin causes the dissociation of both subunits VIa and VIb from the enzyme. These are the two subunits that form the major protein-protein contacts between the two monomeric units within the dimeric complex. Although hydrolysis of CL by phospholipase A(2) and loss of these subunits is linked, the reverse process does not occur, i.e., removal of subunits VIa and VIb does not cause dissociation of the two functionally important, tightly bound cardiolipins. Nor does addition of exogenous cardiolipin result in reassociation of the two subunits with the remainder of the complex. We conclude that cardiolipin is not only essential for full electron transport activity, but also has an important structural role in stabilizing the association of subunits VIa and VIb within the remainder of the bovine heart enzyme.
Sedlák
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Department of Biochemistry, The University of Texas Health Science Center, San Antonio, Texas 78229-3900, USA.
Robinson
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NC
D000818:Animals; D002308:Cardiolipins / Q000378:metabolism*; D002417:Cattle; D017839:Crotalus; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D013329:Structure-Activity Relationship
D002308:Cardiolipins; D003576:Electron Transport Complex IV; D010741:Phospholipases A