Vertiev Iu V, Skvortsov V T, Ezepchuk Iu V
Biokhimiia. 1977 Aug;42(8):1424-31.
A model for studying the interaction of diphtheria toxin with cell membranes includes immobilization of purified cell membranes on Sephadex G-25, adsorbtion of toxin on the membranes in the presence of protective colloid, and subsequent detection of adsorbed toxin by means of virioimmunoassay. Diphtheria toxin adsorbed rapidly on membranes of both sensitive (HeLa, macrophages) and resistent to tis action cells, but not on stromaof human erythrocytes. The rate of interaction depends on the concentration of toxin and the temperature of incubation. Adsorbed toxin may be eluted by acidic buffer, 8 M area and 4 M guanidin. HCl, but not by triton X-100, tween 20 and 80, sodium dodecylaulfate and basic buffer.
将纯化的细胞膜固定在葡聚糖凝胶G - 25上,在保护胶体存在的情况下使毒素吸附在膜上,随后通过病毒免疫测定法检测吸附的毒素。白喉毒素能迅速吸附在敏感细胞(HeLa细胞、巨噬细胞)和对其作用有抗性的细胞的膜上,但不能吸附在人红细胞的基质上。相互作用的速率取决于毒素的浓度和孵育温度。吸附的毒素可用酸性缓冲液、8M脲和4M盐酸胍洗脱,但不能用曲拉通X - 100、吐温20和80、十二烷基硫酸钠和碱性缓冲液洗脱。