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Analysis of Polypeptide and Protein Structures Using Fourier Transform Infrared Spectroscopy

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Microscopy, Optical Spectroscopy, and Macroscopic Techniques

Part of the book series: Methods in Molecular Biology ((MIMB,volume 22))

Abstract

Infrared spectroscopy was one of the earliest techniques to be used for the structural studies of polypeptides and proteins (1,2). However, a major difficulty that limited earlier studies of such biological molecules was the absorption of liquid H2O, which shows strong absorption over much of the fundamental region of the infrared spectrum. This severely limited the analysis of biological molecules in their native state, and necessitated the use of dry films, KBr disks, or D22O as a solvent. There is no strong absorption from D2O, in the region 1700–1500 cm-1and this spectral region is one which is particularly important for the study of polypeptides and proteins. The infrared spectra of liquid H2O and for comparison liquid D2O are shown in Fig. 1. The recent development of computerized FT-IR (Fourier transform infrared) instrumentation now permits the subtraction of background water absorptions from dilute samples (3,4) and, hence, enables the study of biomolecules in their more natural environment, e.g., H2O, buffer solutions, and so on. This approach has revolutionized the application of infrared spectroscopy for the study of biological molecules.

FT-IR transmittance spectrum of H2O (continuous line) and of D2O (broken line) recorded in a calcium fluoride cell fitted with a 6µm tin spacer. The spectra were recorded at 20°C. The peaks shift to lower wave numbers in D2O.

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Authors and Affiliations

  1. Department of Protein and Molecular Biology, Royal Free Hospital School of Medicine, University of London, UK

    Parvez I. Haris & Dennis Chapman

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  1. Parvez I. Haris
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  2. Dennis Chapman
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Editor information

Editors and Affiliations

  1. National Institute for Biological Standards and Control, South Mimms, Potters Bar, UK

    Christopher Jones , Barbara Mulloy  & Adrian H. Thomas ,  & 

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© 1994 Humana Press Inc, Totowa, NJ

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Haris, P.I., Chapman, D. (1994). Analysis of Polypeptide and Protein Structures Using Fourier Transform Infrared Spectroscopy. In: Jones, C., Mulloy, B., Thomas, A.H. (eds) Microscopy, Optical Spectroscopy, and Macroscopic Techniques. Methods in Molecular Biology, vol 22. Humana Press. https://doi.org/10.1385/0-89603-232-9:183

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  • DOI: https://doi.org/10.1385/0-89603-232-9:183

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-232-3

  • Online ISBN: 978-1-59259-509-9

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