Chemical library screening using a SPR-based inhibition in solution assay: simulations and experimental validation.
Anal Chem
; 85(18): 8787-95, 2013 Sep 17.
Article
en En
| MEDLINE
| ID: mdl-23931734
ABSTRACT
We have developed a surface plasmon resonance (SPR)-based inhibition in solution assay (ISA) to search for inhibitors of the medium affinity (KD = 0.8 µM) interaction between an E6-derived peptide (E6peptide) immobilized on the sensor and a PDZ domain (MAGI-1 PDZ1) in the mobile phase. DZ domains are widespread protein-protein interaction modules that recognize the C-terminus of various partners. Simulations indicated that relatively low compound concentrations (10 µM) and limited peptide densities (Rmax < 200 resonance units) should allow the detection of inhibitors with a target affinity close to 100 µM, which was then demonstrated experimentally. ISA screening, carried out on the Prestwick Chemical Library® (1120 compounds), identified 36 compounds that inhibited the interaction by more than 5%. Concentration-dependent ISA, carried out on a subset of 19 potential inhibitors, indicated that 13 of these indeed affected the interaction between MAGI-1 PDZ1 and the E6peptide. No effect was observed for 84 compounds randomly chosen among noninhibitors. One of the four best inhibitors was a peptide binder, and three were PDZ binders with KD in the 10-50 µM range. We propose that a medium (µM) affinity between the target and surface-bound partner is optimal for SPR-based ISA screening.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Simulación por Computador
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Moléculas de Adhesión Celular Neuronal
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Resonancia por Plasmón de Superficie
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Bibliotecas de Moléculas Pequeñas
Tipo de estudio:
Clinical_trials
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Diagnostic_studies
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Screening_studies
Límite:
Humans
Idioma:
En
Revista:
Anal Chem
Año:
2013
Tipo del documento:
Article
País de afiliación:
Francia