Brodner O G, Sabbagh M, Wieland T
Hoppe Seylers Z Physiol Chem. 1982 Mar;363(3):273-8.
The preparation and characterization of a protein from wheat germ showing strong affinity to amatoxins (ABP) but differing from RNA polymerase B (II) is described. The purification, traced by [3H]amatoxin (O-methyldehydroxymethyl-alpha-amanitin), comprises 4 chromatographic steps, on Biogel A 1.5, DEAE-Sephadex, phosphocellulose, and again Biogel A 1.5. The protein exhibited in dodecyl sulfate polyacrylamide gel electrophoresis one single band with a molecular mass of 29,000 Da. Its isoelectric point is 4.9. The dissociation constant of the complex ABP-[3H]amatoxin is KD20 = 5 X 10(-7)M as determined by equilibrium dialysis against alpha-amanitin. By cyanogen bromide the protein is split into two fragments with molecular masses of 22,000 and 7,000 Da, respectively whose amino acid analyses, on summation, give the amino acid composition of ABP.
本文描述了一种从小麦胚芽中提取的蛋白质的制备和特性，该蛋白质对鹅膏毒素具有很强的亲和力（ABP），但与RNA聚合酶B（II）不同。以[3H]鹅膏毒素（O-甲基去氢xymethyl-α-鹅膏蕈碱）追踪的纯化过程包括4个色谱步骤，分别在Biogel A 1.5、DEAE-葡聚糖、磷酸纤维素上进行，然后再次在Biogel A 1.5上进行。该蛋白质在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中显示出一条单一的带，分子量为29,000 Da。其等电点为4.9。通过对α-鹅膏蕈碱进行平衡透析测定，ABP-[3H]鹅膏毒素复合物的解离常数KD20 = 5×10^(-7)M。用溴化氰处理该蛋白质可将其裂解为两个片段，分子量分别为22,000和7,000 Da，将它们的氨基酸分析结果相加，可得出ABP的氨基酸组成。
