RESUMO
The present paper describes the application of step-scan FT-IR spectroscopy in combination with chemometric analysis of the spectral data for the study of the photocycle of bacteriorhodopsin. The focus is on the performance of this instrumentation for time-resolved experiments. Three-dimensional data-spectra recorded over time-are studied using various factor analysis techniques, e.g., singular values decomposition, evolving factor analysis, and multivariate curve resolution based on alternating least squares. Transient intermediates formed in the time domain ranging from 1 micros to 6.6 ms are clearly detected through reliable pure time evolving profiles. At the same time, pure difference absorbance spectra are provided. As a result, valuable information about transitions and dynamics of the protein can be extracted. We conclude first that step-scan FT-IR spectroscopy is a useful technique for the direct study of difficult photochemical systems. Second, and this is the essential motivation of this paper, chemometrics provide a step forward in the description of the photointermediates.
RESUMO
Visible Raman and infrared microspectrometry studies performed on fluorapatite and hydroxyapatite powders have shown similar results. Small modifications of the nu2 and nu4 PO(3-)4 tetrahedra bending modes are observed. A small frequency shift of the nu1 mode and modifications on the nu3 mode region accompanied with a simplification of the hydroxyapatite and fluorapatite respective spectra from seven to four bands were observed. A broad and weak band which could be attributed to the Ca-F bond is detected at 311 cm(-1) on the Raman fluorapatite spectra. The phosphate vibration modes are little disturbed by fluoride substitution. This could indicate that phosphate groups interact strongly between themselves and weakly with substituted atoms (i.e. hydroxyle and fluoride atoms). Whatever crystallographic model is considered, the number of bands observed is always lower than the number of calculated ones, even for hydroxyapatite, whose symmetry is lower than that of fluorapatite.
RESUMO
Elastin is the macromolecular polymer of tropoelastin molecules responsible for the elastic properties of tissues. The understanding of its specific elasticity is uncertain because its structure is still unknown. Here, we report the first experimental quantitative determination of bovine elastin secondary structures as well as those of its corresponding soluble kappa-elastin. Using circular dichroism and Fourier transform infrared and near infrared Fourier transform Raman spectroscopic data, we estimated the secondary structure contents of elastin to be approximately 10% alpha-helices, approximately 45% beta-sheets, and approximately 45% undefined conformations. These values were very close to those we had previously determined for the free monomeric tropoelastin molecule, suggesting thus that elastin would be constituted of a closely packed assembly of globular beta structural class tropoelastin molecules cross-linked to form the elastic network (liquid drop model of elastin architecture). The presence of a strong hydration shell is demonstrated for elastin, and its possible contribution to elasticity is discussed.