Kinetics and protein subunit interactions of Escherichia coli phosphatidylserine decarboxylase in detergent solution.
Phosphatidylserine decarboxylase from Escherichia coli, an intrinsic membrane protein, catalyzes the conversion of phosphatidylserine to phosphatidylethanolamine. The physical and kinetic properties of the purified enzyme were studied in several detergents under assay conditions. The active form of the enzyme is an oligomer, probably a trimer, and the enzyme activity was unaffected by the concentration of the nonionic poly(oxyethylene) ether detergent present in the assay medium, so long as the detergent micelle/substrate mole ratio was less than one. When this ratio was greater than one, the detergent acted as an inhibitor by competing with enzyme-containing micelles for substrate. The zwitterionic and bile salt detergents that were tested inactivated the enzyme by dissociating the oligomer. The native, Triton X-100 solubilized, enzyme was modified with a cross-linking reagent. Activity of the cross-linked enzyme was retained after the Triton X-100 was replaced by a zwitterionic sulfobetaine detergent and conformed to the same kinetic model as with the poly(oxyethylene) ether detergents. The cross-linked enzyme was also active when solubilized by the bile salt detergents although the activity did not conform to any simple kinetic model. These data indicate that the oligomer is the active form of the enzyme under assay conditions and that certain nondenaturing detergents can inactivate this enzyme by dissociating the enzyme complex.
D000818:Animals; D002262:Carboxy-Lyases; D002417:Cattle; D003840:Deoxycholic Acid; D003902:Detergents; D004220:Disulfides; D004926:Escherichia coli / Q000201:enzymology*; D017830:Octoxynol; D010718:Phosphatidylserines; D011092:Polyethylene Glycols; D012967:Sodium Dodecyl Sulfate; D013388:Succinimides; D013501:Surface-Active Agents
D003902:Detergents; D004220:Disulfides; D010718:Phosphatidylserines; D013388:Succinimides; D013501:Surface-Active Agents; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; D002262:Carboxy-Lyases; C022353:phosphatidylserine decarboxylase; C011240:dithiobis(succinimidylpropionate)