Schmidt Miranda L, Davis James H
University of Guelph, Department of Physics, 50 Stone Road E, Guelph, Ontario N1G 2W1, Canada.
Biochim Biophys Acta. 2016 Apr;1858(4):619-26. doi: 10.1016/j.bbamem.2015.12.016. Epub 2015 Dec 17.
Magnetically orienting bicelles are often made by combining the long chain phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) with the short chain phospholipid 1,2-dicaproyl-sn-glycero-3-phosphocholine (DCPC) in buffer. These bicelles orient with their bilayer normals perpendicular to the external magnetic field. We have examined the phase behaviour of DMPC/DCPC bicelles and the effects of cholesterol and the unsaturated phospholipid 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (DPoPC) as a function of temperature using static solid state (2)H nuclear magnetic resonance spectroscopy. As expected, cholesterol has an ordering effect on the long phospholipid chains and this is reflected in the phase behaviour of the bicelle mixtures. Liquid disordered-liquid ordered, fluid-fluid phase coexistence is observed in DMPC/cholesterol/DCPC bicelles with cholesterol mole fractions of 0.13 and higher. DPoPC/DMPC/cholesterol/DCPC bicelles also exhibit two fluid phase coexistence over a broad range of temperatures and compositions. Bicelles can provide a useful medium in which to study membrane bound peptides and proteins. The orientation parallel to the magnetic field is favourable for studying membrane peptides/proteins because information about the orientation of relevant molecular bonds or internuclear vectors can be obtained directly from the resulting (2)H spectra. Lanthanide ions can be used to flip the bicelles to have their bilayer normals parallel to the external magnetic field. Yb(3+) was used to flip the DPoPC/DMPC/cholesterol/DCPC bicelles while Eu(3+) was found to be ineffective at flipping bicelles containing cholesterol in the present work.
磁性定向双分子层通常是通过在缓冲液中将长链磷脂1,2 - 二肉豆蔻酰 - sn - 甘油 - 3 - 磷酸胆碱(DMPC)与短链磷脂1,2 - 二己酰 - sn - 甘油 - 3 - 磷酸胆碱(DCPC)混合制成的。这些双分子层的双层法线方向垂直于外部磁场。我们使用静态固态(2)H核磁共振光谱研究了DMPC/DCPC双分子层的相行为以及胆固醇和不饱和磷脂1,2 - 二棕榈油酰 - sn - 甘油 - 3 - 磷酸胆碱(DPoPC)随温度的影响。正如预期的那样,胆固醇对长磷脂链具有有序化作用,这在双分子层混合物的相行为中得到体现。在胆固醇摩尔分数为0.13及更高的DMPC/胆固醇/DCPC双分子层中观察到液相无序 - 液相有序、流体 - 流体相共存。DPoPC/DMPC/胆固醇/DCPC双分子层在广泛的温度和组成范围内也表现出两种流体相共存。双分子层可以提供一个有用的介质来研究膜结合肽和蛋白质。平行于磁场的取向有利于研究膜肽/蛋白质,因为可以直接从所得的(2)H光谱中获得有关相关分子键或核间矢量取向的信息。镧系离子可用于翻转双分子层,使其双层法线平行于外部磁场。在本工作中,Yb(3+)用于翻转DPoPC/DMPC/胆固醇/DCPC双分子层,而发现Eu(3+)在翻转含胆固醇的双分子层时无效。