Purification and characterization of extracellular phospholipase A2 from human synovial fluid in rheumatoid arthritis.
Extracellular phospholipase A2 was purified about 1.7 X 10(5) fold to near homogeneity from human synovial fluid of rheumatoid arthritis by sequential use of column chromatographies on heparin-Sepharose, butyl-Toyopearl, and reversed-phase HPLC. The final preparation showed a single band on SDS-polyacrylamide gel electrophoresis, and its molecular mass was estimated to be approximately 13,700 daltons. The purified enzyme had a pH optimum of 9.0 and required Ca2+ for maximum activity. It hydrolyzed phosphatidyl-ethanolamine more effectively than phosphatidylserine and phosphatidylcholine. These properties were similar to those of an extracellular phospholipase A2 detected in the peritoneal cavity of caseinate-treated rats.
Hara
S
S
Department of Health Chemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Kudo
I
I
Chang
H W
HW
Matsuta
K
K
Miyamoto
T
T
Inoue
K
K
D001172:Arthritis, Rheumatoid / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D054467:Phospholipases A2; D013379:Substrate Specificity; D013582:Synovial Fluid / Q000201:enzymology*
D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium