Artículo
How to distinguish ligand-binding mechanisms: an example of conformational selection disguised as an induced fit
Fecha de publicación:
10/2019
Editorial:
Inst Biology
Revista:
Journal Of Biological Education
ISSN:
0021-9266
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
This report describes the implementation of a laboratory exercise for an advanced biochemistry or enzyme kinetics class at the undergraduate or graduate level, designed to improve understanding of protein conformational changes associated with the binding of a ligand. Students measure the fluorescence changes induced by the conformational transition of a glycoprotein (the Na,K-ATPase) upon addition of different ligands (Pi and BeF3 −) and analyse the results in order to determine the mechanism of the process. The results show that Pi and BeF3 − present opposite effects on the observed rate constants (kobs) with ligand concentration: kobs decreases with [Pi] and increases with [BeF3 −]. This observation, together with the frequently used assumption that binding occurs under rapid equilibrium, led to propose different models for ligand-induced conformational transitions: a conformational selection for Pi and an induced fit for BeF3 −. In this paper, we show that if the rapid-equilibrium approximation for ligand binding is not assumed, a conformational selection mechanism can account for the effects of both ligands. This active-learning exercise serves as the basis for discussing the consequences of not being extremely cautious when invoking approximations about not-very-well-known systems and the importance of a correct understanding of models assigned to chemical processes.
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Articulos(IQUIFIB)
Articulos de INST.DE QUIMICA Y FISICO-QUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Articulos de INST.DE QUIMICA Y FISICO-QUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Citación
Faraj, Santiago Enrique; Rossi, Rolando Carlos; Lopez Montes, Mónica Beatriz; How to distinguish ligand-binding mechanisms: an example of conformational selection disguised as an induced fit; Inst Biology; Journal Of Biological Education; 10-2019; 1-17
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