Baldacini O, Green G A, Girardot R, Rihn B, Monteil H
Institut de Bactériologie, Faculté de Médecine, Strasbourg, France.
J Chromatogr. 1990 Jun 29;528(2):357-69. doi: 10.1016/s0378-4347(00)82394-4.
We have developed a rapid method for the purification of proteins, combining titration curve analysis with a two-step column chromatographic procedure. We have used this approach to purify the cytotoxin (L toxin) from Clostridium sordellii. We have also determined the amino acid composition of this cytotoxin. This toxin has a pI value of 4.20 and an Mr of 260,000, reduction of which results in a band of Mr 43,000 on sodium dodecyl sulphate polyacrylamide gel electrophoresis. Since both the proteins of Mr 260,000 and 43,000 are recognized by the polyclonal anti-C. sordellii L toxin, which neutralizes the L toxin cytotoxicity, we propose a hexameric structure for the protein of Mr 260,000, each subunit being Mr 43,000.
我们开发了一种蛋白质快速纯化方法,将滴定曲线分析与两步柱色谱法相结合。我们已用此方法从索氏梭菌中纯化细胞毒素(L毒素)。我们还测定了这种细胞毒素的氨基酸组成。该毒素的pI值为4.20,Mr为260,000,将其还原后在十二烷基硫酸钠聚丙烯酰胺凝胶电泳上产生一条Mr为43,000的条带。由于Mr为260,000和43,000的两种蛋白质都能被中和L毒素细胞毒性的多克隆抗索氏梭菌L毒素识别,我们提出Mr为260,000的蛋白质具有六聚体结构,每个亚基的Mr为43,000。