In:
Applied Spectroscopy, SAGE Publications, Vol. 48, No. 11 ( 1994-11), p. 1432-1441
Kurzfassung:
A Fourier transform infrared spectroscopic method has been developed to analyze protein secondary structure by employing the amide III spectral region (1350–1200 cm −1 )· Benefits of using the amide III region have been shown to be substantial. The interference from the water vibration (∼1640 cm −1 ) in the amide I region can be avoided when one is using the amide III band; furthermore, the amide III region also presents a more characterized spectral feature which provides easily resolved and better defined bands for quantitative analysis. Estimates of secondary structure are accomplished with the use of Fourier self-deconvolution, second derivatization, and curve-fitting on original protein spectra. The secondary structure frequency windows (α-helix, 1328–1289 cm −1 ; unordered, 1288–1256 cm −1 ; and β-sheets, 1255–1224 cm −1 ) have been obtained, and estimates of secondary structural contents are consistent with X-ray crystallography data for model proteins and parallel results obtained with the use of the amide I region. We have further applied the analysis to the structural change of calsequestrin upon Ca 2+ binding. Treatment of calsequestrin with 1 mM Ca 2+ results in the formation of crystalline aggregates accompanied by a 10% increase in α-helical structure, which is consistent with previous results obtained by Raman spectroscopy. Thus the amide III region of protein IR spectra appears to be a valuable tool in estimating individual protein secondary structural contents.
Materialart:
Online-Ressource
ISSN:
0003-7028
,
1943-3530
DOI:
10.1366/0003702944028065
Sprache:
Englisch
Verlag:
SAGE Publications
Publikationsdatum:
1994
ZDB Id:
1474251-2
SSG:
11
