Cysteine conformation and sulfhydryl interactions in proteins and viruses. 3. Quantitative measurement of the Raman S-H band intensity and frequency.
The bond stretching vibration of the cysteine sulfhydryl (SH) group in a typical protein generates a Raman band in the spectral interval 2500-2600 cm-1, a region devoid of interference from any other fundamental mode of vibration of the protein. The relatively high Raman cross section associated with the S-H stretching vibration, the sensitivity of the vibrational frequency to hydrogen bonding interactions and side chain configurations, and the dependence of the Raman intensity on thiol-thiolate equilibria, combine to make the Raman SH band a potentially valuable marker of protein sulfhydryl interactions and a unique indicator of sulfhydryl participation in thiol-disulfide oxidoreductase activity. In order to exploit Raman spectroscopy for these purposes, accurate and precise measurements of Raman SH band profiles are required. We show here that the required precision and accuracy can be achieved by use of the Raman band corresponding to the stretching vibration of in situ nitrogen gas as a quantitative intensity and frequency standard. The Raman Q-branch center of the N2 band occurs at 2330.7 cm-1.
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Division of Cell Biology and Biophysics, School of Biological Sciences, University of Missouri-Kansas City 64110.
Vohník
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Thomas
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D017100:Bacteriophage P22 / Q000737:chemistry; D055592:Biophysical Phenomena; D001703:Biophysics; D003545:Cysteine / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry; D008623:Mercaptoethanol; D009584:Nitrogen / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D013059:Spectrum Analysis, Raman; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D015678:Viral Structural Proteins / Q000737:chemistry; D014780:Viruses / Q000737:chemistry*
D004279:DNA, Viral; D011506:Proteins; D013438:Sulfhydryl Compounds; D015678:Viral Structural Proteins; D008623:Mercaptoethanol; D003545:Cysteine; D009584:Nitrogen