Chen-Marotel J, Blouquit Y, Rosa R, Calvin M C
J Chromatogr. 1983 Mar 18;258:213-22. doi: 10.1016/s0021-9673(00)96414-8.
A new method for the purification of human erythrocyte phosphoglycerate-kinase involving affinity chromatography on dye-ligand media (Red A), in the presence of 3-phosphoglycerate and ATP, is described. The method is rapid and technically simple. The purity of the enzyme was verified by electrophoresis in polyacrylamide gel in the presence of sodium dodecylsulphate, by amino acid analysis and by immunoprecipitation in Ouchterlony plates. Peptide mapping of tryptic digests of the purified enzyme was performed and the immunoneutralization of the enzyme activity evaluated with rabbit antibodies.
本文描述了一种在3-磷酸甘油酸和ATP存在的情况下，利用染料配体介质（红色A）亲和色谱法纯化人红细胞磷酸甘油酸激酶的新方法。该方法快速且技术简单。通过在十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳、氨基酸分析以及在Ouchterlony平板上的免疫沉淀法验证了酶的纯度。对纯化酶的胰蛋白酶消化产物进行了肽图谱分析，并用兔抗体评估了酶活性的免疫中和作用。
