Wada A, Tachibana H, Hayashi H, Saito Y
J Biochem Biophys Methods. 1980 May;2(5):257-69. doi: 10.1016/0165-022x(80)90050-0.
A computerized spectrophotometer system which is capable of simultaneously obtaining three spectral dimensions, that is, the absorbance, the circular dichroism and the fluorescence intensity, from one sample solution has been developed for the purpose of attaining a higher resolving power in the study of conformational transitions of biological macromolecules. Measurement conditions, such as the wavelength, the temperature, pH or the concentration of reagents in the sample solution, can be scanned according to a sequence that is set just prior to the measurement. A computer-driven micro-injector and a pH electrode directly immersed in the sample solution make it possible to obtain a titration curve in parallel to the optical measurement. All the data taken are stored on a magnetic disk for later retrieval. They can be processed and displayed in any required form. The helix-coil transitions of a polynucleotide caused by temperature and those of a polypeptide caused by pH, and the denaturation of proteins caused by guanidine hydrochloride, were studied by this measuring system. The continuous plotting of transition profiles and the correlation diagrams among different spectral dimensions has proved to be a good way of demonstrating the existence of different modes of transition.
为了在生物大分子构象转变的研究中获得更高的分辨能力，已开发出一种计算机化分光光度计系统，该系统能够从一个样品溶液中同时获得三个光谱维度，即吸光度、圆二色性和荧光强度。测量条件，如波长、温度、pH值或样品溶液中试剂的浓度，可以根据测量前设定的序列进行扫描。计算机驱动的微量注射器和直接浸入样品溶液中的pH电极使得能够在进行光学测量的同时获得滴定曲线。采集的所有数据都存储在磁盘上以供以后检索。它们可以以任何所需的形式进行处理和显示。利用该测量系统研究了温度引起的多核苷酸的螺旋-卷曲转变、pH值引起的多肽的螺旋-卷曲转变以及盐酸胍引起的蛋白质变性。连续绘制转变曲线以及不同光谱维度之间的相关图已被证明是证明不同转变模式存在的好方法。
