Zalman L S, Müller-Eberhard H J
Research Institute of Scripps Clinic, Department of Immunology, La Jolla, CA 92037.
Mol Immunol. 1990 Jun;27(6):533-7. doi: 10.1016/0161-5890(90)90072-8.
The channels formed by poly C9, C5b-8 and C5b-9 were examined using the liposome swelling assay. By plotting the relative rate of swelling of C5b-8-containing liposomes vs the molecular weight of the sugar solute and by applying the Renkin equation, the size of the C5b-8 channel was estimated to be 1.5 mm radius. As increasing amounts of C9 were added during the formation of C5b-9, in C8:C9 ratios of 1:1, 1:2, 1:6 and 1:12, the size of the function channel increased. Poly C9 had a pore that was somewhat larger than C5b-9 at a C8:C9 ratio of 1:12. Using molecular sieving experiments with four different iodinated protein size markers, the channel diameter of poly C9 was estimated at between 90 and 100 A. Monoclonal antibodies to different complement proteins were added to the liposomes to see which might inhibit the channels. C5b-8 containing liposomes could be inhibited by antibodies to C8. Liposomes containing C5b-9 could be inhibited slightly by antibodies to C9 and most strongly by antibodies to the neoantigen of poly C9.
使用脂质体肿胀试验检测了由多聚C9、C5b-8和C5b-9形成的通道。通过绘制含C5b-8脂质体的相对肿胀率与糖溶质分子量的关系图,并应用伦金方程,估计C5b-8通道的半径为1.5毫米。在形成C5b-9的过程中,随着添加的C9量增加,C8:C9比例为1:1、1:2、1:6和1:12时,功能性通道的尺寸增大。在C8:C9比例为1:12时,多聚C9的孔径比C5b-9略大。使用四种不同的碘化蛋白质大小标记物进行分子筛实验,估计多聚C9的通道直径在90至100埃之间。将针对不同补体蛋白的单克隆抗体添加到脂质体中,以观察哪些抗体可能抑制通道。含C5b-8的脂质体可被抗C8抗体抑制。含C5b-9的脂质体可被抗C9抗体轻微抑制,而被抗多聚C9新抗原的抗体抑制作用最强。