Polymorphic phase behavior of unsaturated lysophosphatidylethanolamines: a 31P NMR and X-ray diffraction study.
Polymorphic phase behavior of unsaturated lysophosphatidylethanolamines: a 31P NMR and X-ray diffraction study.
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不饱和溶血磷脂酰乙醇胺的多晶型相行为：31P NMR 和 X 射线衍射研究。
DOI：
10.1021/bi00352a013
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Gruner,SM
中科院分区：
文献类型：
--
作者：
Tilcock,CP;Cullis,PR;Hope,MJ;Gruner,SM
材料和方法脂质合成。如前所述，合成并提纯了二油酰磷脂酰乙醇胺、二亚油酰基-PE和二芳烯基-PE(Tilcock&Cullis，1982)。Lyso-PE由它们各自的双酰基PE通过磷脂酶A2水解得到，如下所示。通常，将1g双酰基PE溶解到100毫升对数级乙醚中，其中添加20毫升0.5M三(羟甲基)氨基甲烷盐酸盐(Tris-HCl)(pH 7.4)、20毫升2 mM CaCl2和50毫克Crotalusadamanteus蛇毒(Sigma)。反应容器用氮气冲洗并密封，混合物在室温下黑暗中大力搅拌。24-36h后，薄层层析[CHCl3/MeOH/25%NH3/H20(90：54：5.7：5.3v/v)；硅胶60预涂层板，Merck]证明完全转化为Lyso衍生物。用旋转蒸发法脱除大部分乙醚，用4体积的CHCl3/MeOH(2：1v/v)提取3次水相。萃取物被混合在一起，浓缩成油。采用Waters Prep-500柱，硅胶为载体，CHCl3/MeOH/H20(60：30：3v/v)为洗脱液，用制备液层析纯化Lyso-PE(-gt；98%)。最后通过CM-柱层析实现了最终纯化。
Materials and MethodsLipid Synthesis. Dioleoylphosphatidylethanolamine, di-linoleoyl-PE, and dilinolenoyl-PE were synthesized and purified as described previously (Tilcock & Cullis, 1982). Lyso-PE’s were derived from their respective diacyl-PE’s by phospholipase A2 hydrolysis as follows. Typically, 1 g of diacyl-PE was dissolved to clarity in 100 mL of analar-grade diethyl ether to which was added 20 mL of 0.5 M tris (hydroxymethyl)-aminomethane hydrochloride (Tris-HCl)(pH 7.4), 20 mL of 2 mM CaCl2, and 50 mg of Crotalusadamanteus venom (Sigma). The reaction vessel was flushed with nitrogen and sealed, and the mixture was stirred vigorously at room tem-perature in the dark. After 24-36 h, complete conversion to the lyso derivative occurred as evidenced by thin-layer chromatography [CHCl3/MeOH/25% NH3/H20 (90: 54: 5.7: 5.3 v/v); silica gel 60 precoated plates, Merck]. The majority of the ether was removed by rotary evaporationand the aqueous phase extracted 3 times with 4 volumes of CHCl3/MeOH (2: 1 v/v). The extracts were pooled and concentrated to oils. Lyso-PE’s were purified (> 98%) by preparative liquid chromatography using a Waters Prep-500 LC unit with silica as support and CHCl3/MeOH/H20 (60: 30: 3 v/v) as eluant. Final purification was achieved by chromatography on CM-