synchrotron radiation circular dichroism and conventional circular dichroism spectroscopy a comparison同步加速器辐射圆二色性和传统的圆二色性光谱进行比较.pdfVIP

Spectroscopy 16 (2002) 121–125 121 IOS Press Synchrotron radiation circular dichroism and conventional circular dichroism spectroscopy: A comparison J.G. Lees and B.A. Wallace ∗ Department of Crystallography, Birkbeck College, University of London, London WC1E 7HX, UK Abstract. Conventional circular dichroism (cCD) spectroscopy is a valuable tool for secondary structure analyses of proteins. In recent years, it has been possible to use synchrotrons as light sources for CD, with the technique being known as Synchrotron Radiation Circular Dichroism (SRCD). In this study, the spectra of two proteins, the primarily helical myoglobin and the pri- marily beta-sheet concanavalin A, have been collected on both a cCD instrument and on the SRCD at the Daresbury synchrotron and their characteristics were compared. Over the wavelength regions where both instruments are capable of making measure- ments (from about 300 to 175 nm) the spectra are very similar, except at the low wavelength extreme of the cCD spectra. In this region, the spectra deviate somewhat, due to the limitations of the light source intensity in the conventional instrument. The SRCD spectra extend to much lower wavelengths (160 nm). This additional low wavelength vacuum ultraviolet (VUV) data contains a large amount of extra information, including, for the first time, a number of peaks consistent with previously predicted charge transfer transitions. 1. Introduction Circular Dichroism (CD) is a popular technique for determining the secondary structural contents of proteins. With modern conventional circular dichroism (cCD) instruments, data can be collected to as low as ∼175 nm for a protein in aqueous solution. Since the absorption of water increases sharply below wavelengths of around 200 nm, and the flux of cCD instruments decreases dramatically over this same wavelength range, this limits