Immobilized liposome chromatography for studies of protein-membrane interactions and refolding of denatured bovine carbonic anhydrase

摘要: Small unilamellar vesicles (SUVs) composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and 1 mol% phosphatidylethanolamine were covalently coupled to chromatographic gel beads. Interactions liposomal lipid bilayers with several water-soluble proteins, which had been denatured or partially by 0.1-5 M guanidinium hydrochloride (GuHCl), studied on beads containing the immobilized SUVs. The partially-denatured proteins treated 0.5-1.0 GuHCl significantly retarded liposome column, whereas little retardation native unfolded >2 was observed same columns. column found be well correlated local hydrophobicity, determined aqueous two-phase partitioning method using mM Triton X-405 as a hydrophobic probe. It implies that are likely in molten-globule state associated bilayers. Chromatographic refolding bovine carbonic anhydrase (CAB) achieved column. enzymatic activity an CAB 5 recovered up 83% after passing it through only 58% when run liposome-free process is probably involved interaction