Chiang T M, Alkan M L, Beachey E H
Infect Immun. 1979 Oct;26(1):316-21. doi: 10.1128/iai.26.1.316-321.1979.
The spontaneous binding of group A streptococcal lipoteichoic acid (LTA) to mammalian cell membranes was studied in isolated membranes of human erythrocytes. The binding of radiolabeled LTA to erythrocyte membranes was dependent on membrane concentration and time. Binding approached a maximum within 30 min of incubation. The bound LTA could be displaced by adding a 50-fold excess of unlabeled LTA. The displaced LTA was eluted from a column of Sepharose 6B in a position identical to that of authentic LTA, suggesting that binding did not alter the size of the molecule. A dissociation constant of 42 micrometers was calculated, and only one population of approximately 5.5 X 10(6) binding sites per erhtyrocyte membrane was detected. Since these results suggested that erythrocyte membranes possess specific binding sites for LTA, an attempt was made to localize the putative receptors to the outside or the inside surface of the erhtyrocyte membrane. Assays of the binding of LTA to resealed right-side-out and inside-out membrane ghosts demonstrated that the outside surface was able to bind over 10 times more LTA than the inside surface. These results support the concept that the membranes possess specific binding sites for LTA and inciate that these binding sites are located almost entirely on the outside surface of erythrocyte membranes.
在人红细胞的分离膜中研究了A组链球菌脂磷壁酸(LTA)与哺乳动物细胞膜的自发结合。放射性标记的LTA与红细胞膜的结合取决于膜浓度和时间。孵育30分钟内结合接近最大值。加入50倍过量的未标记LTA可取代结合的LTA。被取代的LTA从琼脂糖6B柱上洗脱的位置与 authentic LTA相同,表明结合未改变分子大小。计算出解离常数为42微米,并且在每个红细胞膜上仅检测到约5.5×10(6)个结合位点的一个群体。由于这些结果表明红细胞膜具有LTA的特异性结合位点,因此尝试将推定的受体定位到红细胞膜的外表面或内表面。对LTA与重新封闭的外翻和内翻膜空泡结合的测定表明,外表面结合的LTA比内表面多10倍以上。这些结果支持了膜具有LTA特异性结合位点的概念,并表明这些结合位点几乎完全位于红细胞膜的外表面。