Your browser doesn't support javascript.
loading
Fluorescence Polarization-Based Measurement of Protein-Ligand Interaction in Fungal Cell Lysates.
LeBlanc, Emmanuelle V; Shekhar-Guturja, Tanvi; Whitesell, Luke; Cowen, Leah E.
Afiliación
  • LeBlanc EV; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
  • Shekhar-Guturja T; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
  • Whitesell L; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
  • Cowen LE; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
Curr Protoc ; 1(1): e17, 2021 Jan.
Article en En | MEDLINE | ID: mdl-33484500
Fungi infect over a billion people worldwide and contribute substantially to human morbidity and mortality despite all available therapies. New antifungal drugs are urgently needed. Decades of study have revealed numerous protein targets of potential therapeutic interest for which potent, fungal-selective ligands remain to be discovered and developed. To measure the binding of diverse small molecule ligands to their larger protein targets, fluorescence polarization (FP) can provide a robust, inexpensive approach. The protocols in this article provide detailed guidance for developing FP-based assays capable of measuring binding affinity in whole cell lysates without the need for purification of the target protein. Applications include screening of libraries to identify novel ligands and the definition of structure-activity relationships to aid development of compounds with improved target affinity and fungal selectivity. © 2021 Wiley Periodicals LLC. Basic Protocol 1: Use of saturation binding curves to optimize tracer and lysate protein concentrations Basic Protocol 2: Establishment of competition binding experiments Support Protocol 1: Preparation of fungal cell lysates Support Protocol 2: Preparation of human HepG2 cell lysate.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas / Hongos Tipo de estudio: Guideline Límite: Humans Idioma: En Revista: Curr Protoc Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas / Hongos Tipo de estudio: Guideline Límite: Humans Idioma: En Revista: Curr Protoc Año: 2021 Tipo del documento: Article País de afiliación: Canadá Pais de publicación: Estados Unidos