title: Effect of ethanol-water mixture on the structure and dynamics of lysozyme: a fluorescence correlation spectroscopy study.
abstract: Effect of ethanol-water mixture on the hydrodynamic radius (r(H)) and conformational dynamics of lysozyme has been studied by circular dichroism, emission spectra, and fluorescence correlation spectroscopy. For this purpose, the protein lysozyme is covalently labeled near the active site with a fluorescent probe, alexa 488. The ethanol molecules are sequestered near the hydrophobic tryptophan residues as indicated by the blue shift of the emission maximum of tryptophan. It is observed that both size (r(H)) and time constant of conformational relaxation (τ(R)) of lysozyme oscillate with increase in ethanol concentration. The r(H) of the protein fluctuates from 19 Å in the native state, to a minimum of 13 Å, and a maximum of 29 Å. It is proposed that the oscillating behavior arises from competition between mutual interaction among protein, ethanol, and water. The fluorescence intensity fluctuates because of quenching of the fluorescence of the probe (alexa) by the free amino group of certain residues (e.g., tryptophan). Rate of inter-conversion (folding dynamics) between the open (fluorescent) and closed (non-fluorescent) form has been determined and is found to exhibit similar oscillation with variation in ethanol content.
authors.affiliation: Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata - 700032, India.
authors.forename: Shyamtanu
authors.initials: S
authors.lastname: Chattoraj
authors.affiliation: Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata - 700032, India.
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Mandal
authors.affiliation: Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata - 700032, India.
authors.forename: Kankan
authors.initials: K
authors.lastname: Bhattacharyya
mesh_terms: D000431:Ethanol / Q000737:chemistry*; D057927:Hydrophobic and Hydrophilic Interactions; D015394:Molecular Structure; D009113:Muramidase / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics; D014867:Water / Q000737:chemistry*
chemical_list: D014867:Water; D000431:Ethanol; D009113:Muramidase