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Abstract

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  2. Abstract
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Attenuated total reflection Fourier-transform infrared spectroscopy of thin hydrated films of soluble and membrane protein included in a phospholipid bilayer is shown to provide useful information as to the secondary structure of the protein. The analysis of the amide I band of deuterated samples by Fourier self-deconvolution followed by a curve fitting was performed by a new procedure in which all the input parameters are generated by the computer rather than by the investigator. The results of this analysis provide a correct estimation of the α-helix and β-sheet structure content with a standard deviation of 8.6% when X-ray structures are taken as a reference. We also show that the orientation of the different secondary structures resolved by the Fourier self-deconvolution/curve-fitting procedure and of the phospholipid acyl chains can be simultaneously evaluated for membrane proteins reconstituted in a lipid bilayer. Of special interest for reconstitution of membrane proteins, the lipid/protein ratio can be accurately and quickly determined from the infrared spectrum.

Abbreviations
ATR

attenuated total reflection

FTIR

Fourier-transform infrared spectroscopy

Myr2GroPCho

dimyristoylphos-phatidylcholine

OcGlc

octyl-glucopyranoside

FWHH

full width at half height

REFERENCES

  1. Top of page
  2. Abstract
  3. REFERENCES