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Print ISSN : 0919-2719
Regular Article
Chemometric Methodology for the Isolation of Absorption Spectrum of Dye Probe Associated with Biological Macromolecules
Yoshiaki Nakamaru Chusuke Sato
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ジャーナル フリー

1999 年 7 巻 2 号 p. 79-88

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Many dye probes have been widely used in morphological fields, but their mechanisms as probes remain unknown in almost all cases. We have explored a possible methodology for discussing the physiological states of the probes bound to biological macromolecules, and found that factor analysis (FA), one of the methods of multivariate analysis, is useful for isolating the absorption spectrum of a dye probe associated with protein molecules. As an example of the simplified reaction system for protein-ligand interactions, bovine serum albumin (BSA) and bromocresol purple (BCP) were used, and the basic methodology was developed using the FA calculations. In principle, there are two types of data series in standard spectrophotometry: spectral series obtained with various concentrations of the protein at constant amounts of the dye probe (Type-I data), and those obtained with various concentrations of the dye probe at constant amounts of the protein (Type-II data). These data types have different data matrices. The data type depends on the experimental conditions, but the Type-II data are more usual than the Type-I data. Therefore, it is important to confirm the applicability of the FA calculations to both data types. In this study we focused on FA calculations using mainly Type-II data and confirmed the same results as with Type-I data. Possible application of this methodology to in vivo observation is also discussed.
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