Role of Ca2+ in the binding of phospholipase A2 with a monomeric substrate and with its amide-type analog.
Effects of Ca2+ on the kinetic parameters for the hydrolysis of monodispersed 1,2-dihexanoyl-sn-glycero-3-phosphorylcholine (diC6PC), catalyzed by Group I phospholipases A2 (PLA2s) from Pseudechis australis, Naja naja atra, and bovine pancreas and by Group II enzymes from Vipera russelli russelli, Agkistrodon halys blomhoffii, and Trimeresurus flavoviridis, were studied by the pH-stat assay method at 25 degrees C, pH 7.5-8.2, and an ionic strength of 0.1 or 0.2 in the absence or presence of an amide-type substrate analog, 2-dodecanoyl-amino-1-hexanol-phosphoglycol. The binding of genuine substrate to the Group II enzymes and that of its analog to the Groups I and II enzymes were markedly facilitated by the binding of Ca2+ to the enzymes. On the other hand, the binding of genuine substrate to the Group I enzymes was found to be independent of the Ca2+ binding. The former result suggests that the structures of the Group II enzyme-genuine substrate complexes and both types of enzyme-analog complexes are generally stabilized by the Ca2+ binding, whereas the latter indicates that the structures of the Group I enzyme-genuine substrate complexes are already similar to those of their Ca2+ complexes and that, therefore, these enzyme-substrate interactions are independent of the Ca2+ binding.
Fujii
S
S
Department of Biochemistry, Osaka University of Pharmaceutical Sciences.
Tani
T
T
Hada
S
S
Inoue
S
S
Ikeda
K
K
Iwama
S
S
Katsumura
S
S
Samejima
Y
Y
Omori-Satoh
T
T
Takasaki
C
C
D000577:Amides / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002417:Cattle; D004546:Elapid Venoms / Q000201:enzymology; D006868:Hydrolysis; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D010713:Phosphatidylcholines / Q000378:metabolism*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054467:Phospholipases A2; D014757:Viper Venoms / Q000201:enzymology
C092445:1,2-dihexanoyl-sn-glycero-3-phosphorylcholine; D000577:Amides; D004546:Elapid Venoms; D007527:Isoenzymes; D010713:Phosphatidylcholines; D014757:Viper Venoms; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium