External reflection FTIR of peptide monolayer films in situ at the air/water interface: experimental design, spectra-structure correlations, and effects of hydrogen-deuterium exchange
| Autor: | Joseph W. Brauner, R.C. Baldwin, Carol R. Flach, J.W. Taylor, Richard Mendelsohn |
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| Rok vydání: | 1994 |
| Předmět: |
Infrared
Protein Conformation Molecular Sequence Data Analytical chemistry Biophysics Infrared spectroscopy Protein Structure Secondary chemistry.chemical_compound Structure-Activity Relationship Amide Monolayer Spectroscopy Fourier Transform Infrared Amino Acid Sequence Fourier transform infrared spectroscopy Air Circular Dichroism Water Models Theoretical Deuterium Kinetics chemistry Hydrogen–deuterium exchange Peptides Oligopeptides Water vapor Mathematics Research Article Hydrogen |
| Zdroj: | Biophysical Journal. 67(1):402-410 |
| ISSN: | 0006-3495 |
| DOI: | 10.1016/s0006-3495(94)80495-3 |
| Popis: | A Fourier transform infrared spectrometer has been interfaced with a surface balance and a new external reflection infrared sampling accessory, which permits the acquisition of spectra from protein monolayers in situ at the air/water interface. The accessory, a sample shuttle that permits the collection of spectra in alternating fashion from sample and background troughs, reduces interference from water vapor rotation-vibration bands in the amide I and amide II regions of protein spectra (1520–1690 cm-1) by nearly an order of magnitude. Residual interference from water vapor absorbance ranges from 50 to 200 microabsorbance units. The performance of the device is demonstrated through spectra of synthetic peptides designed to adopt alpha-helical, antiparallel beta-sheet, mixed beta-sheet/beta-turn, and unordered conformations at the air/water interface. The extent of exchange on the surface can be monitored from the relative intensities of the amide II and amide I modes. Hydrogen-deuterium exchange may lower the amide I frequency by as much as 11–12 cm-1 for helical secondary structures. This shifts the vibrational mode into a region normally associated with unordered structures and leads to uncertainties in the application of algorithms commonly used for determination of secondary structure from amide I contours of proteins in D2O solution. |
| Databáze: | OpenAIRE |
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