Inouye H, Karthigasan J, Kirschner D A
Department of Neurology Research, Children's Hospital, Boston, Massachusetts.
Biophys J. 1989 Jul;56(1):129-37. doi: 10.1016/S0006-3495(89)82658-X.
The biochemical composition of myelin and the topology of its constituent lipids and proteins are typically studied using membranes that have been isolated from whole, intact tissue using procedures involving hypotonic shock and sucrose density gradient centrifugation. To what extent, however, are the structure and intermembrane interactions of isolated myelin similar to those of intact myelin? We have previously reported that intact and isolated myelins do not always show identical myelin periods, indicating a difference in membrane-membrane interactions. The present study addresses the possibility that this is due to altered membrane structure. Because x-ray scattering from isolated myelin sometimes consists of overlapping Bragg reflections or is continuous, we developed nonlinear least squares procedures for analyzing the total intensity distribution after film scaling, background subtraction, and Lorentz correction. We calculated electron density profiles of isolated myelin for comparison with membrane profiles from intact myelin. The change in the width of the extracellular space and the relative invariance of the cytoplasmic space as a function of pH and ionic strength that we previously found for intact nerve was largely paralleled by isolated myelin. There were two exceptions: isolated CNS myelin was resistant to swelling under all conditions, and isolated PNS myelin in hypotonic saline showed indefinite swelling at the extracellular apposition. However, electron density profiles of isolated myelins, calculated to 30 A resolution, did not show any major change in structure compared with intact myelin that could account for the differences in interactions.
髓磷脂的生化组成及其组成脂质和蛋白质的拓扑结构通常使用通过涉及低渗休克和蔗糖密度梯度离心的程序从完整的全组织中分离出来的膜进行研究。然而,分离出的髓磷脂的结构和膜间相互作用在多大程度上与完整髓磷脂的相似呢?我们之前报道过,完整的和分离出的髓磷脂并不总是呈现相同的髓鞘周期,这表明膜 - 膜相互作用存在差异。本研究探讨了这可能是由于膜结构改变所致的可能性。由于从分离出的髓磷脂进行的X射线散射有时由重叠的布拉格反射组成或呈连续状,我们开发了非线性最小二乘法程序,用于在薄膜缩放、背景扣除和洛伦兹校正后分析总强度分布。我们计算了分离出的髓磷脂的电子密度分布,以便与完整髓磷脂的膜分布进行比较。我们之前在完整神经中发现的细胞外空间宽度随pH和离子强度的变化以及细胞质空间的相对不变性在很大程度上与分离出的髓磷脂相似。有两个例外情况:分离出的中枢神经系统髓磷脂在所有条件下都抗肿胀,而在低渗盐溶液中的分离出的周围神经系统髓磷脂在细胞外附着处显示出无限期肿胀。然而,计算分辨率达30埃的分离出的髓磷脂的电子密度分布与完整髓磷脂相比,并未显示出任何能解释相互作用差异的结构上的重大变化。