Schengrund C L, Ringler N J, Dasgupta B R
Department of Biological Chemistry, M. S. Hershey Medical Center, Pennsylvania State University, Hershey 17033.
Brain Res Bull. 1992 Dec;29(6):917-24. doi: 10.1016/0361-9230(92)90166-u.
The ability of 125I-labeled botulinum type A and tetanus neurotoxins to adhere to blots of synaptosomal proteins separated by SDS-polyacrylamide gel electrophoresis was studied. Both neurotoxins appeared to adhere preferentially to an approximately 80 kDa and to a lesser extent to an approximately 116 kDa protein(s). Adherence of the neurotoxins to these proteins was enhanced by preincubation of the neurotoxins with GT 1b. The approximately 100 kDa heavy chain segment of BTxA adhered to the same proteins. The carboxy terminal half of the heavy chain adhered primarily to the approximately 80 kDa protein(s) while the amino terminal portion bound most intensely to the approximately 116 kDa protein(s). The ability of the approximately 80 and approximately 116 kDa proteins to stain positively with the periodic acid-Schiff reagent and to bind 125I-labeled wheat germ lectin suggests that they are glycosylated. Both neurotoxins appear to adhere to the same approximately 80 and approximately 116 kDa proteins because tetanus neurotoxin preincubated with GT 1b was able to reduce binding of radiolabeled botulinum type A neurotoxin to both proteins. Neither neurotoxin adhered to blots of proteins from liver, spleen, or kidney, suggesting that the proteins adhered to are neural components.
研究了125I标记的A型肉毒杆菌毒素和破伤风神经毒素与经SDS-聚丙烯酰胺凝胶电泳分离的突触体蛋白印迹的结合能力。两种神经毒素似乎都优先结合一种约80 kDa的蛋白，在较小程度上结合一种约116 kDa的蛋白。神经毒素与GT 1b预孵育可增强其与这些蛋白的结合。BTxA约100 kDa的重链片段与相同的蛋白结合。重链的羧基末端一半主要与约80 kDa的蛋白结合，而氨基末端部分与约116 kDa的蛋白结合最紧密。约80 kDa和约116 kDa的蛋白用高碘酸-希夫试剂呈阳性染色并结合125I标记的麦胚凝集素，表明它们是糖基化的。两种神经毒素似乎都与相同的约80 kDa和约116 kDa的蛋白结合，因为与GT 1b预孵育的破伤风神经毒素能够减少放射性标记的A型肉毒杆菌毒素与这两种蛋白的结合。两种神经毒素均不与肝、脾或肾的蛋白印迹结合，提示所结合的蛋白是神经成分。
