Phospholipase A2 engineering. Structural and functional roles of the highly conserved active site residue aspartate-49.
Phospholipase A2 engineering. Structural and functional roles of the highly conserved active site residue aspartate-49.
复制标题
磷脂酶A2工程。
DOI：
10.1021/bi00253a009
复制
发表时间：
1994
期刊：
影响因子：
2.9
通讯作者：
Tsai,MD
中科院分区：
文献类型：
--
作者：
Li,Y;Yu,BZ;Zhu,H;Jain,MK;Tsai,MD
材料和方法材料和常规程序。寡核苷酸获自Bio-Synthesis（Lewisville，TX），并且不经进一步纯化而使用。诱变和测序试剂盒分别从Bio-Rad和United States Bio-chemicals获得。DCgPC、DC 14 PC、DOPC和POPC购自Avanti Polar Lipids（Birmingham，AL）。如前所述制备本研究中使用的以下脂质：DC 14 PM（Jain & Gelb，1991）;脱氧-LPC和MJ 33（Jain等人，1991 c）; DTPM（Jain等人，1986 a）; MG 14由华盛顿大学（西雅图）的Michael Gelb教授提供。超纯盐酸胍（Gdn-HCl）获自ICN Biochemical; 99.9原子% D2 G、“100%”D2 G和3-（三甲基甲硅烷基）丙酸钠-2，2，3，J-d4（TMSP）购自MSD Isotopes。DC 1购自
MATERIALS AND METHODSMaterials and Routine Procedures. Oligonucleotides were obtained from Bio-Synthesis (Lewisville, TX) and used without further purification. Mutagenesis and sequencing kits were obtained from Bio-Rad and United States Bio-chemicals, respectively. DCgPC, DC14PC, DOPC, and POPC were purchased from Avanti Polar Lipids (Birmingham, AL). The following lipids used in thisstudy were prepared as previously described: DC 14PM (Jain & Gelb, 1991); deoxy-LPC and MJ33 (Jain et al., 1991c); DTPM (Jain et al., 1986a); MG14 was provided by Professor Michael Gelb at the University of Washington (Seattle). Ultrapure guanidine hydrochloride (Gdn-HCl) was obtained from ICN Biochemical; 99.9 atom% D D2G,“100%” D2G, and sodium 3-(trimethylsilyl) propionate-2, 2, 3, J-d4 (TMSP) were pur-chased from MSD Isotopes. DC1 was purchased from