Excimer fluorescence to monitor transitions of the association dissociation equilibria induced by dilution of proteins composed of subunits.
Dilution is a simple method to induce shifts of the association-dissociation equilibria of protein subunits. Excimer fluorescence is useful to monitor changes of equilibria in dilute solution, which has been shown in a well-characterized protein, actin. Actin was labeled with a fluorescent probe; N-(1-pyrenyl)iodoacetamide. When actin was diluted in low ionic strength buffer, the relative excimer fluorescence decreased. This arose from the conformational drift of actin, which resulted in a change in the association-dissociation equilibria of actin subunits in the dilute solution. Since dilution effects are often observed in other proteins, excimer fluorescence would be a useful sensitive technique to detect shifts of association-dissociation equilibria in dilute solution.
Aichi Prefectrural University of Fine Arts, Japan.
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Ikkai
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Kondo
D000199:Actins / Q000737:chemistry; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005453:Fluorescence; D007700:Kinetics; D046911:Macromolecular Substances; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012996:Solutions; D014461:Ultracentrifugation
D000199:Actins; D046911:Macromolecular Substances; D011506:Proteins; D012996:Solutions