title: Protein interactions of Merocyanine 540: spectroscopic and crystallographic studies with lysozyme as a model protein.
abstract: Spectroscopic and crystallographic studies reveal that Merocyanine 540 (MC 540), a well-known therapeutically important anionic cyanine dye, interacts with hen egg white lysozyme in ground state. The formation of the complex is validated by two isosbestic points in absorption spectra of lysozyme with varied concentration of MC 540 and appearance of an isodichroic point in induced CD spectra of MC 540 with lysozyme. The blue shift of fluorescence maximum of lysozyme in presence of MC 540 shows hydrophobic effect on Trp due to complex formation probably through cooperative binding. Above 1:3M stoichiometric ratio (lysozyme:MC 540) an additional fluorescence hump arises because of structural changes in protein, where MC 540 acts as self-denaturant, inducing non-linearity in Stern-Volmer plot. The van't Hoff isotherms with negative changes in enthalpy at lower concentration and positive changes in entropy for entire concentration range of MC 540 depict the binding forces as hydrogen bonding/van der Waal's and ionic/hydrophobic respectively. Finally X-ray crystallographic structure of the complex shows that MC 540 adopts two conformations, cis and trans, while it binds to lysozyme. Benzoxole moiety of MC 540 interacts with Trp123 through π-stacking and SO3(2-) group is stabilized by ionic interaction/H-bonding with Arg125 of lysozyme.
authors.affiliation: Chemical Sciences Division, Saha Institute of Nuclear Physics, 1/AF, Bidhannagar, Kolkata 700 064, India.
authors.forename: Piyali
authors.initials: P
authors.lastname: Mitra
authors.forename: Mousumi
authors.initials: M
authors.lastname: Banerjee
authors.forename: Sampa
authors.initials: S
authors.lastname: Biswas
authors.forename: Samita
authors.initials: S
authors.lastname: Basu
mesh_terms: D000818:Animals; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D009113:Muramidase / Q000737:chemistry*; D011485:Protein Binding; D011744:Pyrimidinones / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
chemical_list: D011744:Pyrimidinones; C003954:merocyanine dye; C421864:hen egg lysozyme; D009113:Muramidase