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Probing Ligand–Protein Recognition with Sum‐Frequency Generation Spectroscopy: The Avidin–Biocytin Case
Author(s) -
Dreesen Laurent,
Sartenaer Yannick,
Humbert Christophe,
Mani Alaa A.,
Méthivier Christophe,
Pradier ClaireMarie,
Thiry Paul A.,
Peremans André
Publication year - 2004
Publication title -
chemphyschem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.016
H-Index - 140
eISSN - 1439-7641
pISSN - 1439-4235
DOI - 10.1002/cphc.200400213
Subject(s) - biocytin , avidin , chemistry , sum frequency generation spectroscopy , spectroscopy , ligand (biochemistry) , sum frequency generation , biotin , biochemistry , physics , biology , optics , neuroscience , receptor , quantum mechanics , central nervous system , laser , nonlinear optics
Infrared/visible sum‐frequency generation (SFG) spectroscopy is used to study the recognition of a protein (avidin) by a derived vitamin (biocytin) adsorbed on a calcium fluoride substrate. The specificity of the process is tested by replacing avidin with bovine serum albumin or presaturated avidin. The SFG spectroscopy shows drastic modifications in the CH and NH spectral ranges only upon exposure of the biocytin film to avidin. The comparison of the SFG data with Fourier transform infrared reflection absorption spectra (FT‐IRRAS) in the same spectral ranges illustrates the advantages of nonlinear spectroscopy for studying and detecting recognition between biomolecules.