Donta S T, Poindexter N J, Ginsberg B H
Biochemistry. 1982 Feb 16;21(4):660-4. doi: 10.1021/bi00533a011.
The binding of iodinated cholera and Escherichia coli (LT) enterotoxins to Y1 mouse adrenal cells was studied by using saturation analysis (Scatchard). Each toxin bound to Y1 cells with similar affinity [KA = (1.5--2.0) x 10(9)M-1], but there appeared to be twice as many receptor sites per cell for E. coli toxin (approximately 4 x 10(5). Despite the increased binding of E. coli toxin, Y1 cells respond sooner to, and to smaller concentrations of, cholera toxin. The binding of each toxin was inhibited competitively by both toxins, although twice as much E. coli toxin was required to inhibit 50% of the binding of cholera toxin as was needed for either homologous inhibition or the inhibition of E. coli toxin binding by cholera toxin. The B subunits of both toxins were equally effective in competing for the binding of both iodinated toxins. Whereas the A subunits of both toxins had little or no effect on the binding of E. coli toxin, they consistently inhibited 20--40% of the binding of cholera toxin to cells. These results suggest that there are receptor loci on cells for the A subunit and that conformational differences exist between the two toxins that might explain the greater sensitivity of Y1 cells to cholera toxin. A model is suggested in which cholera toxin exhibits a greater degree of multivalent ligand binding than does the E coli toxin, resulting in a more favorable situation for apposition of the A subunit to its receptor or for its insertion into the membrane.
采用饱和分析(Scatchard法)研究了碘化霍乱毒素和大肠杆菌(LT)肠毒素与Y1小鼠肾上腺细胞的结合情况。每种毒素与Y1细胞的结合亲和力相似[平衡解离常数KA = (1.5 - 2.0)×10⁹ M⁻¹],但大肠杆菌毒素每个细胞的受体位点似乎是霍乱毒素的两倍(约4×10⁵个)。尽管大肠杆菌毒素的结合量增加,但Y1细胞对霍乱毒素的反应更快,且所需浓度更低。两种毒素都能竞争性抑制对方的结合,不过抑制50%霍乱毒素结合所需的大肠杆菌毒素量是同源抑制或霍乱毒素抑制大肠杆菌毒素结合所需量的两倍。两种毒素的B亚基在竞争结合两种碘化毒素方面同样有效。两种毒素的A亚基对大肠杆菌毒素的结合几乎没有影响,但它们始终能抑制20% - 40%的霍乱毒素与细胞的结合。这些结果表明细胞上存在A亚基的受体位点,且两种毒素之间存在构象差异,这可能解释了Y1细胞对霍乱毒素更敏感的原因。提出了一个模型,其中霍乱毒素比大肠杆菌毒素表现出更高程度的多价配体结合,从而为A亚基与受体的并置或其插入膜中创造了更有利的条件。