title: Microviscosity-induced conformational transition in β-lactoglobulin in the presence of an ionic liquid.
abstract: This study reports on the helix-beta conformation transition of bovine β-lactoglobulin (βLG) prepared at two different pH conditions (pH 4 and 7.5) and in the presence of the ionic liquid 1-ethyl-3-methylimidazolium ethyl sulfate (IL-emes). The investigation was carried out by combining a range of techniques such as circular dichroic (CD) spectroscopy, steady-state fluorescence spectroscopy, isothermal titration calorimetry (ITC), and transmission electron microscopy. The influence of microviscosity induced by IL-emes on the secondary structure of βLG was studied using a quartz crystal microbalance and correlated with the steady-state fluorescence emission. The effect of heat on the helix-beta transition in βLG was directly measured by ITC by titrating βLG with IL-emes. The net effect of heat after subtraction of the heat of dilution was negative in both cases, suggesting that the protein moves to a stable conformation. The changes in the overall aggregated structures were confirmed by transmission electron microscopy, where a shift in the size and morphology of aggregates was found, from large clusters (size of 70 nm) at pH 4 to smaller aggregates (size of 20 nm) at pH 7.5, which reduced to 7 nm in the presence of the IL. The transformation of helical to beta structure at pH 4 show that the folding pathway in the presence of the ionic liquid is hierarchical, whereas at neutral pH, it appeared to be nonhierarchical and the final native structure was acquired by nonlocal interactions through typical forces involved in the stabilization of the tertiary structure.
authors: Chemical Laboratory, CSIR-CLRI, Adyar, Chennai 600020, India.
authors: Kamatchi
authors: K
authors: Sankaranarayanan
authors: B
authors: B
authors: Sreedhar
authors: B U
authors: BU
authors: Nair
authors: A
authors: A
authors: Dhathathreyan
mesh_terms: D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D052578:Ionic Liquids / Q000494:pharmacology*; D007782:Lactoglobulins / Q000737:chemistry*; D008958:Models, Molecular; D011487:Protein Conformation / Q000187:drug effects; D014783:Viscosity
chemical_list: D052578:Ionic Liquids; D007782:Lactoglobulins