Conformational transitions in beta-lactoglobulin induced by cationic amphiphiles: equilibrium studies.
The conformational transition from the native state in water ("beta-state") to a state containing a considerable amount of alpha-helices ("alpha-state") was studied for the protein beta-lactoglobulin (BLG), from bovine milk, in several colloidal solutions containing mixed micelles or spontaneous vesicles. These aggregates were formed in the bicationic system containing the surfactant dodecyltrimethylammonium chloride (DTAC) and the lipid didodecyldimethylammonium bromide (DDAB). The beta-->alpha transition in BLG, investigated by far-ultraviolet circular dichroism spectroscopy, is induced to the same protein alpha-state by pure and mixed DDAB/DTAC micelles or vesicles. This implies a similar interaction mechanism of BLG with DDAB or DTAC, once the colloidal aggregates are formed. In premicelle DTAC solutions, the fraction of alpha-helix is lower and increases with the DTAC concentration. DDAB and DTAC also promote conformational changes in the protein tertiary structure that expose the tryptophans to a less constrained environment. These unfolding transitions were investigated by near-ultraviolet circular dichroism and steady-state fluorescence spectroscopies. In equilibrium conditions, it was found that higher DTAC (and, probably, DDAB) concentrations are needed to induce the beta-->alpha transition than to unfold the protein. beta-Lactoglobulin may therefore be considered as a model for protein-surfactant and protein-lipid interactions.
Viseu
Maria Isabel
MI
Centro de Química Estrutural, Instituto Superior Técnico, 1049-001 Lisbon, Portugal. iviseu@popsrv.ist.utl.pt
Carvalho
Teresa Isabel
TI
Costa
Sílvia M B
SM
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism / Q000379:methods; D007782:Lactoglobulins / Q000737:chemistry*; D008823:Micelles; D008894:Milk Proteins / Q000737:chemistry; D008958:Models, Molecular; D011489:Protein Denaturation; D017510:Protein Folding; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry
D007782:Lactoglobulins; D008823:Micelles; D008894:Milk Proteins; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; C046112:didodecyldimethylammonium; C022806:dodecyltrimethylammonium