McMullen T P, Wong B C, Tham E L, Lewis R N, McElhaney R N
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Biochemistry. 1996 Dec 24;35(51):16789-98. doi: 10.1021/bi962098t.
It has been proposed that the lower levels of exogenous cholesterol incorporation into the membranes of the sterol-non-requiring as compared to the sterol-requiring mycoplasmas may be due to the much higher glycolipid content of the former and to the reduced ability of glycolipids, as opposed to phospholipids, to incorporate sterols [Efrati et al. (1986) Arch. Biochem. Biophys. 248, 282-288]. In order to test this hypothesis, we have investigated the interaction of cholesterol with the major membrane glyco- and phospholipids of the sterol-non-requiring mycoplasma Acholeplasma laidlawii B, utilizing elaidic acid-homogenous membranes in order to obviate any differences in the nature of cholesterol-lipid interactions due to variations in the fatty acid composition of the different membrane components. Specifically, we have studied the effect of increasing quantities of cholesterol on the thermotropic phase behavior of aqueous dispersions of phosphatidylglycerol, diglucosyl diacylglycerol, and monoglucosyl diacylglycerol, as well as the total membrane polar lipids of this organism, using high-sensitivity differential scanning calorimetry. We find that cholesterol is highly miscible in both the lamellar gel and liquid-crystalline states of phosphatidylglycerol but exhibits limited miscibility in the two neutral glycolipids, particularly in their lamellar gel and crystalline states. We also demonstrate that cholesterol has a limited miscibility in both the lamellar gel and liquid-crystalline states of bilayers composed of the total A. laidlawii B membrane polar lipids. These results demonstrate that the nature of cholesterol-lipid interactions depends markedly on the structure of the glycerolipid polar headgroup and suggests that the incorporation of lower levels of cholesterol into the membranes of the sterol-non-requiring mycoplasmas may indeed be due, at least in part, to their high glycolipid contents. We also show that cholesterol stabilizes the lamellar liquid-crystalline phase of the monoglucosyl diacylglycerol relative to the inverted hexagonal phase at all sterol concentrations, in contrast to the effects of cholesterol on dielaidoylphosphatidylethanolamine, which destabilizes the lamellar liquid-crystalline phase at low concentrations.
有人提出,与需要固醇的支原体相比,不需要固醇的支原体膜中外源胆固醇掺入水平较低,可能是由于前者的糖脂含量高得多,而且与磷脂相比,糖脂掺入固醇的能力降低[埃弗拉蒂等人(1986年),《生物化学与生物物理学档案》248卷,282 - 288页]。为了验证这一假设,我们研究了胆固醇与不需要固醇的支原体莱氏无胆甾原体B的主要膜糖脂和磷脂的相互作用,使用反油酸均一膜以消除由于不同膜成分脂肪酸组成变化而导致的胆固醇 - 脂质相互作用性质的任何差异。具体而言,我们使用高灵敏度差示扫描量热法研究了增加量的胆固醇对磷脂酰甘油、二葡糖基二酰基甘油和单葡糖基二酰基甘油以及该生物体总膜极性脂质的水分散体热致相行为的影响。我们发现胆固醇在磷脂酰甘油的层状凝胶态和液晶态中高度互溶,但在两种中性糖脂中互溶性有限,特别是在它们的层状凝胶态和结晶态中。我们还证明胆固醇在由莱氏无胆甾原体B总膜极性脂质组成的双层膜的层状凝胶态和液晶态中互溶性有限。这些结果表明胆固醇 - 脂质相互作用的性质明显取决于甘油olipid极性头部基团的结构,并表明在不需要固醇的支原体膜中掺入较低水平的胆固醇可能确实至少部分归因于它们高糖脂含量。我们还表明,与胆固醇对二反油酸磷脂酰乙醇胺的影响相反,在所有固醇浓度下,胆固醇相对于反相六角相稳定了单葡糖基二酰基甘油的层状液晶相,后者在低浓度下会破坏层状液晶相。