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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
- Source :
- Biophysical Journal. 67(1):402-410
- Publication Year :
- 1994
- Publisher :
- Elsevier BV, 1994.
-
Abstract
- 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.
- Subjects :
- 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
Subjects
Details
- ISSN :
- 00063495
- Volume :
- 67
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Biophysical Journal
- Accession number :
- edsair.doi.dedup.....29350ce13d43ea0b73020348478f8ff0
- Full Text :
- https://doi.org/10.1016/s0006-3495(94)80495-3