The chemical step is not rate-limiting during the hydrolysis by phospholipase A2 of mixed micelles of phospholipid and detergent.
The chemical step is not rate-limiting during the hydrolysis by phospholipase A2 of mixed micelles of phospholipid and detergent.
复制标题
在磷脂酶A2水解磷脂和洗涤剂的混合胶束的过程中，化学步骤不是限速的。
DOI：
10.1021/bi00083a040
复制
发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Berg,OG
中科院分区：
文献类型：
--
作者：
Jain,MK;Rogers,J;Hendrickson,HS;Berg,OG
材料与方法按照描述合成smj33和deoxy-LPC (Jain et al., 1991d)， DMPM和POPM (Jain & Gelb, 1991)，以及二硫代dmpc （Hendrickson & Hendrickson, 1990）。本研究中使用的其他磷脂均来自Avanti。胆汁盐和CHAPSO来自Sigma， Triton X-100来自boehringer。从猪胰腺和毒液中分离出PLA2 （Jain et al., 1991b）。底物磷脂（通常为20 mg/mL）在水中的原液分散体在浴式超声器中通过超声制备。用pH-stat滴定法监测反应过程（Upreti & Jain, 1978, 1980； Jain等人，1986a； Berg等人，1991）。反应混合物中含有4ml在pH 8.0和23 C下剧烈搅拌（转速2000 rpm，不起泡）的溶液，其中含有1mm NaCl和1mm（阴离子磷脂）或6mm CaCh（两性离子磷脂）。基质浓度保持不变
MATERIALS AND METHODSMJ33 and deoxy-LPC (Jain et al., 1991d), DMPM and POPM (Jain & Gelb, 1991), and dithio-DMPC (Hendrickson & Hendrickson, 1990) were synthesized as described. Other phospholipids used in this study were from Avanti. Bile salts and CHAPSO were from Sigma, and Triton X-100 was fromBoehringer. PLA2 from pig pancreas and venoms were isolated as described (Jain et al., 1991b). Stock dispersions of substrate phospholipids (typically 20 mg/mL) in water were prepared by sonication in a bath-type sonicator. The reaction progress was monitored by pH-stat titration (Upreti & Jain, 1978, 1980; Jain et al., 1986a; Berg et al., 1991). The reaction mixture contained 4 mL of vigorously stirred (at 2000 rpm without frothing) solution at pH 8.0 and 23 C containing 1 mM NaCl and 1 mM (for anionic phospholipids) or 6 mM CaCh (for zwitterionic phospholipids). The bulk substrate concentration was kept