title: Obtaining information about protein secondary structures in aqueous solution using Fourier transform IR spectroscopy.
abstract: Fourier transform IR (FTIR) spectroscopy is a nondestructive technique for structural characterization of proteins and polypeptides. The IR spectral data of polymers are usually interpreted in terms of the vibrations of a structural repeat. The repeat units in proteins give rise to nine characteristic IR absorption bands (amides A, B and I-VII). Amide I bands (1,700-1,600 cm(-1)) are the most prominent and sensitive vibrational bands of the protein backbone, and they relate to protein secondary structural components. In this protocol, we have detailed the principles that underlie the determination of protein secondary structure by FTIR spectroscopy, as well as the basic steps involved in protein sample preparation, instrument operation, FTIR spectra collection and spectra analysis in order to estimate protein secondary-structural components in aqueous (both H2O and deuterium oxide (D2O)) solution using algorithms, such as second-derivative, deconvolution and curve fitting. Small amounts of high-purity (>95%) proteins at high concentrations (>3 mg ml(-1)) are needed in this protocol; typically, the procedure can be completed in 1-2 d.
authors: Department of Chemistry, Fudan University, Shanghai, China.
authors: Huayan
authors: H
authors: Yang
authors: Department of Chemistry, Fudan University, Shanghai, China.
authors: Shouning
authors: S
authors: Yang
authors: Department of Chemistry, Fudan University, Shanghai, China.
authors: Jilie
authors: J
authors: Kong
authors: Department of Chemistry and Biochemistry, University of Northern Colorado, Greeley, Colorado, USA.
authors: Aichun
authors: A
authors: Dong
authors: Department of Chemistry, Fudan University, Shanghai, China.
authors: Shaoning
authors: S
authors: Yu
mesh_terms: D053000:Analytic Sample Preparation Methods / Q000379:methods*; D002124:Calcium Fluoride / Q000737:chemistry; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D017666:Deuterium Oxide / Q000737:chemistry; D017433:Protein Structure, Secondary / Q000235:genetics*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water; D017666:Deuterium Oxide; D002124:Calcium Fluoride
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