Measuring and Analyzing Binding Kinetics of Coupled Folding and Binding Reactions Under Pseudo-First-Order Conditions.

Jensen, Kristine Steen

Jensen, Kristine Steen.

Afiliación

Jensen KS; Department for Biophysical Chemistry, Center for Molecular Protein Science, LTH, Lund University, Lund, Sweden. kristine.steen_jensen@bpc.lu.se.

Methods Mol Biol ; 2141: 629-650, 2020.

Article en En | MEDLINE | ID: mdl-32696381

RESUMEN

Many intrinsically disordered proteins (IDPs) adopt a well-defined structure upon binding to their interaction partners. Kinetic characterization is a requirement for the investigation of the dynamics and mechanisms of these folding-upon-binding reactions. Here a protocol is described for the investigation of binding kinetics of bimolecular binding and folding reactions of IDPs to their ligand partner under pseudo-first-order conditions using stopped-flow mixing and fluorescence detection.

Asunto(s)

Proteínas Intrínsecamente Desordenadas/metabolismo; Unión Proteica; Pliegue de Proteína; Diseño de Equipo; Proteínas Intrínsecamente Desordenadas/química; Cinética; Ligandos; Modelos Moleculares; Conformación Proteica; Espectrometría de Fluorescencia/métodos; Termodinámica

Palabras clave

Association and dissociation rate constants; Binding mechanisms; Displacement kinetics; Folding and binding; IDP; Intrinsically disordered proteins; Stopped-flow measurements

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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Unión Proteica / Pliegue de Proteína / Proteínas Intrínsecamente Desordenadas Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2020 Tipo del documento: Article País de afiliación: Suecia Pais de publicación: Estados Unidos

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