SINGLE-STEP PURIFICATION OF RAT-LIVER ALDOLASE USING IMMOBILIZED ARTIFICIAL MEMBRANE CHROMATOGRAPHY

Aldolase B is a peripheral membrane protein. Immobilized artificial me mbrane (IAM) surfaces were used to purify rat liver aldolase B in a si ngle chromatographic step. Selective elution required dipalmitoylphosp hatidylcholine (DPPC) to be included in the mobile phase. Selective el ution of aldolase from the IAM column when DPPC (0.2 mM) was added to the mobile phase indicates that DPPC was an affinity displacing ligand for this membrane associated protein. Since tissue preparation involv ed only homogenization and centrifugation, the single step purificatio n of aldolase B using IAM chromatography is a very convenient method. The IAM stationary phase (1.5 g) has a loading capacity of at least 4. 39 mg total protein from rat liver homopenates and typically approxima te to 17.7 mu g of pure aldolase in a single step from approximate to 60 mg wet weight rat liver cytosol can be obtained. (C) 1997 Elsevier Science B.V.