Lectin-carbohydrate complex evaluation by chemiluminescence.
Anal Biochem
; 548: 91-95, 2018 05 01.
Article
en En
| MEDLINE
| ID: mdl-29458035
ABSTRACT
In order to characterize the affinity between specific carbohydrate-binding proteins such as lectins, a model is proposed to study these interactions using a polysaccharide membrane to simulate such adsorption. Here, lectin-carbohydrate interactions were chemiluminescently investigated using lectins conjugated to acridinium ester (AE) and polysaccharides composed of their respective specific carbohydrates. The lectin-AE conjugates were incubated with discs (0.0314-0.6358â¯cm2) of phytagel, chitosan and carrageenan. The complex formation chemiluminescently detected followed the Langmuir isotherm from which constants were estimated. The association constant (Ka) and maximum binding sites on the membranes were 2.4â¯×â¯10-7â¯M-1⯱â¯0.8â¯×â¯10-7â¯M-1 and 1.3â¯×â¯10-3â¯mol. mg-1 ± 0.3â¯×â¯10-3â¯mol. mg-1 (Con A); 0.9â¯×â¯10-6â¯M-1⯱â¯0.4â¯×â¯10-6â¯M-1 and 0.021â¯×â¯10-3â¯mol. mg-1 ± 0.003â¯×â¯10-3â¯mol. mg-1 (WGA) and 2.0â¯×â¯10-6â¯M-1⯱â¯0.9â¯×â¯10-6â¯M-1 and 0.069â¯×â¯10-3â¯mol. mg-1 ± 0.010â¯×â¯10-3â¯mol. mg-1 (PNA). The proposed model might be useful to study binding affinity and estimate the amount of binding not limited by the sugar content in the membrane.
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Banco de datos:
MEDLINE
Asunto principal:
Lectinas de Plantas
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Chondrus
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Quitosano
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Mediciones Luminiscentes
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Membranas Artificiales
Idioma:
En
Año:
2018
Tipo del documento:
Article