Effects of relative band intensity on prediction of protein secondary structure from CD

Increasing the magnitude of a protein CD spectrum obviously increases the magnitude of each predicted secondary structure by the same amount. However, increasing the magnitude of the negative, long-wave-length portion of a protein CD spectrum usually has the opposite effect from increasing the positive, short-wave-length portion. Thus small distortions in the CD spectra of proteins at short wavelength can have a significant effect on the analysis for secondary structure. This measurement error and its effect on the analysis are systematically investigated for 16 proteins of known structure. The results demonstrate that a two-point calibration of a CD instrument is mandatory to avoid serious errors when estimating secondary structure from protein CD spectra.