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It should be stressed at this point that due to different polarization conditions the ATR technique is more sensitive to changes in the amide I modes of α-helixes than transmission spectroscopy (cf. ref 15).
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41 published O-BR difference spectra of E204 mutants. The close agreement of their spectra with those presented here (Figure 4) confirms our findings. Minor differences may be explained by using different techniques (ATR, transmission) and different measuring conditions (pH, temperature, time) and by the lower signal-to-noise ratio of the difference spectra of Kandori et al.
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