사람의 중성구 elastase 와 cathepsin G 의 순수분리와 성질

• Published in: BMB reports Vol. 22; no. 4; pp. 379 - 387
• Main Authors: 강구일, 김사열, 정혜영, 배성준, Koo Il Kang, Sa Youl Chim, Hye Young Joung, Sung Jun Bae
• Format: Journal Article
• Language: Korean
• Published: 생화학분자생물학회 01.01.1989
• ISSN: 1976-6696

Human neutrophil elastase(HNE) and human neutrophil cathepsin G (HNCG) were purified by a two-step procedure involving gel filtration through Ultrogel AcA54 and ion-exchange chromatography through CM-sephadex C-25. The purified elastase and cathepsin G cross-reacted with anti-HNE antibody and anti-HNCG antibody respectively. Three elastases have molecular weights of 29,000, 30,000, and 30,500 determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Two cathepsin Gs have the molecular weights of 28,500 and 29,000. There was another group of elastases which showed molecular weights of 2000 dalton higher than the other group of elastase. The discrepancy or the diversity of molecular weights of HNE seems to be caused mainly by the disparity of intra-molecular disulfide bonds of HNE. Monovalent ions including Na^+, Li^+, K^+ and Cs^+ stimulated HNE by concentration dependency. Divalent ions also stimulated HNE very effectively at the concentration of less than 40 mM and then reached the plateau. HNE was completely inhibited by less than 1 mM of diisopropyl fluorophosphate (DIFP), phenylmethylsulfonyl fluoride (PMSF), α₁-protease inhibitor (α₁-PI), and α₂-macroglobulin (α₂-MG) which are catagorized as general endoprotease inhibitor. But leupeptin, which is known as serine and thiol-protease inhibitor, was ineffective on inhibition of HNE.