title: Equimolar mixture of 2,2,2-trifluoroethanol and 4-chloro-1-butanol is a stronger inducer of molten globule state: isothermal titration calorimetric and spectroscopic studies.
abstract: A mixture of 4-chloro-1-butanol and 2,2,2-Trifluoroethanol (TFE) has been used to generate Molten globule (MG) state of structurally homologous but functionally different proteins bovine alpha-lactalbumin and hen egg-white lysozyme. The thermal denaturation was done using UV-Visible spectroscopy. From UV-Visible profile, thermal transition was not observed beyond a particular concentration. There was an indication of molten globule state in case of alpha-lactalbumin from circular dichroism experiments. By intrinsic tryptophan fluorescence, acrylamide and potassium iodide quenching, 8-anilino-naphthalene sulfonic acid (ANS) binding and energy transfer studies the presence of molten globule state was confirmed. Quantitative characterization of MG state and determining the binding thermodynamics of ANS to the MG state was done using Isothermal Titration Calorimetry (ITC). Results show that alpha-lactalbumin exists in MG state at a particular concentration but lysozyme does not show features of MG state.
authors.affiliation: Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai, 400 076, India.
authors.forename: Anu A
authors.initials: AA
authors.lastname: Thoppil
authors.forename: Nand
authors.initials: N
authors.lastname: Kishore
mesh_terms: D000817:Anilino Naphthalenesulfonates / Q000737:chemistry; D000818:Animals; D000440:Butanols / Q000737:chemistry; D002151:Calorimetry; D002417:Cattle; D002724:Chlorobutanol / Q000031:analogs & derivatives* / Q000737:chemistry; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D007768:Lactalbumin / Q000737:chemistry; D009113:Muramidase / Q000737:chemistry; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013050:Spectrometry, Fluorescence; D013696:Temperature; D013816:Thermodynamics; D014270:Trifluoroethanol / Q000737:chemistry*