Wallach D F, Kamat V B, Gail M H
J Cell Biol. 1966 Sep;30(3):601-21. doi: 10.1083/jcb.30.3.601.
Specific turbidities, densities, and refractive indices of fragments of plasma membrane (PM) and endoplasmic reticulum (ER) from Ehrlich ascites carcinoma have been measured. A spherical shell model of specified dimensions and refractive index was established for PM fragments. The ionic composition of the dispersion medium was varied systematically. Increases in Gamma/2 caused increases in the turbidity of both PM and ER suspensions, the greatest effects being observed with Ca(2+) and Mg(2+). In the case of PM this effect is attributable mainly to aggregation, whereas "structural" changes account for most of the turbidity increase with ER. The pH was also varied systematically to obtain pH- density and turbidity profiles and to establish the isoelectric pH of the two membrane types (PM-3.6; ER-4.35). Turbidity was maximum at "isoelectric" pH, which corresponds in each case to the region of minimum charge on the particle surfaces. Both PM and ER show large increases of density at the "isoelectric" pH, but only ER shows substantial structurally based turbidity increase under these conditions. Both PM and ER show operation of electrostatic attractions near "isoelectric" pH. PM has been shown to have ionically distinctive inner and outer surfaces while ER shows no such dissymmetry. The necessary theoretical background for interpretation of turbidity and density measurements is included, as well as a discussion of the limitations of our conclusions and the biological importance of our results.
已对艾氏腹水癌细胞质膜(PM)和内质网(ER)片段的特定浊度、密度和折射率进行了测量。为PM片段建立了具有特定尺寸和折射率的球壳模型。系统地改变了分散介质的离子组成。γ/2的增加导致PM和ER悬浮液的浊度增加,其中Ca(2+)和Mg(2+)的影响最大。就PM而言,这种影响主要归因于聚集,而“结构”变化是ER浊度增加的主要原因。还系统地改变pH值以获得pH-密度和浊度曲线,并确定两种膜类型的等电点pH值(PM - 3.6;ER - 4.35)。在“等电”pH值下浊度最大,在每种情况下,该pH值对应于颗粒表面电荷最小的区域。在“等电”pH值下,PM和ER的密度都大幅增加,但只有ER在这些条件下表现出基于结构的显著浊度增加。在“等电”pH值附近,PM和ER都表现出静电吸引作用。已证明PM具有离子特性不同的内表面和外表面,而ER则没有这种不对称性。文中包含了解释浊度和密度测量结果所需的理论背景,以及对我们结论局限性的讨论和结果的生物学意义。