Continuous titration of difference absorption upon binding of 4-methylumbelliferyl glycosides to concanavalin A and peanut agglutinin
| Autor: | Hilde De Boeck, Frank G. Loontiens, Clement K. De Bruyne |
|---|---|
| Rok vydání: | 1982 |
| Předmět: |
Peanut agglutinin
Arachis Biophysics Analytical chemistry Disaccharide Biochemistry chemistry.chemical_compound Peanut Agglutinin Structure-Activity Relationship Lectins Concanavalin A Glycosides Umbelliferones Molecular Biology Chromatography biology Lectin Cell Biology Chromophore Cuvette chemistry biology.protein Titration Spectrophotometry Ultraviolet Plant Lectins Saturation (chemistry) Hymecromone Protein Binding |
| Zdroj: | Analytical biochemistry. 124(2) |
| ISSN: | 0003-2697 |
| Popis: | A continuous titration of absorption differences is described. Equal volumes of the titration fluid are dispensed from two micrometer-driven Hamilton gas-tight syringes into two 1 × 1 × 4.5-cm cuvettes. These are placed in the reference and sample beam. Each cuvette stopper is equipped with a capillary inlet connected to a syringe and with a minimotor for continuous stirring. Details of the stirring device are given. The delivered volumes of titration fluid are sufficiently reproducible to allow titration of absorption differences as a function of chromophore concentration. The usefulness of this approach is tested with the binding of 4-methylumbelliferyl α- d -mannopyranoside and concanavalin A as a well-characterized system. It is applied to the binding of similarly labeled anti- t disaccharide with the lectin from peanuts. With both lectins, the change in molecular extinction coefficient of the ligand and the association constant, valid for the entire protein saturation range, were obtained. The results are identical to those from other methods, including equilibrium dialysis. |
| Databáze: | OpenAIRE |
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