Partly folded states of bovine carbonic anhydrase interact with zwitterionic and anionic lipid membranes.
The acidic, partly folded states of bovine carbonic anhydrase II (BCAII) were used as an experimental system to study the interactions of partly denatured proteins with lipid membranes. The pH dependence of their interactions with palmitoyloleoyl phosphatidylcholine (POPC) and palmitoyloleoyl phosphatidylglycerol (POPG) membranes was studied. A filtration binding assay shows that acidic partly folded states of BCAII bind to POPC membranes. Fluorescence emission spectra from Trp residues of the bound protein are slightly shifted to shorter wavelength and can be quenched by a water-soluble quencher of fluorescence, indicating that the binding occurs without deep penetration of Trp residues into the membrane. The content of beta-structures of the protein in solution, as revealed by FT-IR spectroscopy, decreases in the partly folded states and the binding to POPC membrane occurs without further changes of secondary structure. In the presence of 0.1 M NaCl, a partly folded state self-aggregates and does not bind to POPC membrane. At acidic pH, BCAII binds to POPG membranes both at high and low ionic strength. The binding to the anionic lipid occurs with protein self-aggregation within the lipid-protein complexes and with changes in the secondary structure; large blue shifts in the fluorescence emission spectra and the decrease in the exposure to water-soluble acrylamide quencher of Trp fluorescence strongly suggest that BCAII penetrates the hydrocarbon domain in the POPG-protein complexes.
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Centro de Investigaciones en Química Biológica de Córdoba, Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Pabellón Argentina, Ciudad Universitaria, 5000, Córdoba, Argentina.
D020106:Acrylamide; D000818:Animals; D000838:Anions / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry*; D002417:Cattle; D002850:Chromatography, Gel; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D009994:Osmolar Concentration; D010713:Phosphatidylcholines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared
D000838:Anions; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D020106:Acrylamide; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; D002256:Carbonic Anhydrases; C028694:1-palmitoyl-2-oleoylphosphatidylcholine