Isolation and fundamental properties of a phospholipase A2 inhibitor from the blood plasma of Trimeresurus flavoviridis.
Phospholipase A2 inhibitor was purified from the blood plasma of Habu, Trimeresurus flavoviridis, by Sephadex G-200 gel filtration, DEAE-cellulose chromatography, and Blue-Sepharose CL-6B column chromatography. The purified inhibitor was shown to be a glycoprotein with a molecular weight of about 100K. It was found to consist of four subunits whose molecular weights were around 20-24K. In order to examine the inhibition mechanism of the inhibitor, the interaction of the inhibitor with a phospholipase A2 from T. flavoviridis venom was examined by Sephadex G-100 gel filtration. One inhibitor molecule was found to bind directly to one phospholipase A2 molecule in both the presence and absence of Ca2+. The inhibitor inhibited the phospholipase A2 from T. flavoviridis venom with an apparent dissociation constant, Ki, of 1.7 X 10(-10) M, but not the porcine pancreas enzyme or the Agkistrodon halys blomhoffii enzyme belonging to the same family, Crotalidae, as T. flavoviridis, or the phospholipase C from Bacillus cereus.
Kogaki
H
H
Department of Biology, Faculty of Science, Osaka University.
Inoue
S
S
Ikeda
K
K
Samejima
Y
Y
Omori-Satoh
T
T
Hamaguchi
K
K
D000818:Animals; D002850:Chromatography, Gel; D004305:Dose-Response Relationship, Drug; D006023:Glycoproteins / Q000097:blood / Q000302:isolation & purification*; D007700:Kinetics; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000097:blood / Q000302:isolation & purification; D054467:Phospholipases A2; D012685:Sepharose / Q000031:analogs & derivatives; D012911:Snakes / Q000097:blood*
D006023:Glycoproteins; C029483:Blue-Sepharose; D012685:Sepharose; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2