The Preparation of Ligandin with Glutathione‐S‐Transferase Activity from Porcine Liver Cytosol by Affinity Chromatography on Bromosulphophthalein‐Sepharose

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Abstract
A simple and rapid method for the purification of porcine ligandin with glutathione-S-transferase activity is presented. After ion-exchange chromatography on DEAE-Sephadex, ligandin is isolated from porcine liver cytosol by affinity chromatography on bromosulfophthalein-Sepharose and gel filtration on Sephadex G-100. Evidence is presented that the purified ligandin is homogeneous with respect to polyacrylamide-gel electrophoresis (7.5%) and sodium dodecylsulfate-gel electrophoresis. Physico-chemical investigations show that the purified ligandin has properties similar to those of ligandin isolated from other species with respect to MW, amino-acid composition, secondary structure and catalytic activity. As is the case for human and rat ligandin, porcine ligandin binds bilirubin. Evidence is also presented that porcine liver cytosol contains several bromosulfophthalein-binding proteins with basic isoelectric points lacking catalytic activity.