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Induced chirality upon binding of cis ‐parinaric acid to bovine β‐lactoglobulin: spectroscopic characterization of the complex
Author(s) -
Zsila Ferenc,
Imre Tı́mea,
Szabó Pál T.,
Bikádi Zsolt,
Simonyi Miklós
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)02771-0
Subject(s) - chemistry , circular dichroism , monomer , crystallography , absorption spectroscopy , chromophore , bovine serum albumin , chirality (physics) , analytical chemistry (journal) , absorption (acoustics) , photochemistry , chromatography , organic chemistry , polymer , materials science , physics , chiral symmetry breaking , quantum mechanics , nambu–jona lasinio model , composite material , quark
Binding of the polyunsaturated cis ‐parinaric acid to bovine β‐lactoglobulin (BLG) was studied by circular dichroism (CD), electronic absorption spectroscopy and mass spectrometry methods. Upon protein binding, the UV absorption band of parinaric acid is red shifted by ca. 5 nm, showing hypochromism and reduced vibrational fine structure, suggesting that the ligand binds as a monomer in non‐planar geometry. In the CD spectra measured at pH 7.36 and 8.5 a strong, negative Cotton band appears centered at 310 nm (Δ ϵ =−25 M −1 cm −1 ) corresponding to the long‐wavelength absorption band of cis ‐parinaric acid. The source of this induced optical activity is the helical distortion of the polyene chromophore caused by the chiral protein environment. From CD spectral data the value of the association constant was calculated to be 4.7×10 5 M −1 at pH 7.36. CD and mass spectrometry measurements showed that parinaric acid binds weakly to BLG in acidic solution, though small peaks at mass 18559 and 18645 can be obtained in the reconstructed electrospray mass spectrum; these correspond to the binding of parinaric acid in 1:1 stoichiometry to both monomer variants of BLG B and A. The hydrophobic interior cavity of BLG was assigned as the primary binding site of cis ‐parinaric acid.