Identification of Ligand Binding by Protein Stabilization: Comparison of ATLAS with Biophysical and Enzymatic Methods
| العنوان: | Identification of Ligand Binding by Protein Stabilization: Comparison of ATLAS with Biophysical and Enzymatic Methods |
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| المؤلفون: | Bin Li, Lindsay J. Howett, Shaohui Wang, Rupal Patel, Richard E. Showalter, Mei-Mei Wang, April Averill, Peggy A. Thompson, James R. Appleman |
| المصدر: | ASSAY and Drug Development Technologies. 6:69-81 |
| بيانات النشر: | Mary Ann Liebert Inc, 2008. |
| سنة النشر: | 2008 |
| مصطلحات موضوعية: | Protein Denaturation, Hot Temperature, Light, Protein Conformation, Biophysics, Drug Evaluation, Preclinical, Calorimetry, Ligands, Biophysical Phenomena, Adenosine Triphosphate, Genes, Reporter, Drug Discovery, Scattering, Radiation, Histidine, Luciferase, Enzyme Inhibitors, Luciferases, chemistry.chemical_classification, Dose-Response Relationship, Drug, Atlas (topology), Circular Dichroism, Ligand binding assay, Proteins, Ligand (biochemistry), Kinetics, Enzyme, chemistry, Biochemistry, Homogeneous, Calibration, Molecular Medicine, Indicators and Reagents, Target protein, Protein stabilization, Oligopeptides |
| الوصف: | ATLAS (Any Target Ligand Affinity Screen) (Anadys Pharmaceuticals, Inc., San Diego, CA) is a homogeneous, affinity-based high-throughput screening technology based on protein thermal denaturation and the ability of ligands to bind and stabilize the target protein from unfolding. To further understand the assay sensitivity for the identification of ligands that bind to soluble protein targets, firefly luciferase was chosen to characterize the technology. Luciferase is a multidomain protein with a complex unfolding pathway. Binding of ATP results in a stabilizing conformational rearrangement of the domains. Using luciferase to characterize the ATLAS technology allowed us to evaluate the generality of the screening method for the identification of ligand binding to any target. Luciferase inhibitors identified from functional screens were used to assess the capability of ATLAS to rank order inhibitors. Comparison of the ATLAS 50% effective concentration with other biophysical and biochemical methods offered insight into optimizing ATLAS assay conditions to maximize sensitivity to compound binding and protein stabilization. The results show the importance of characterizing the thermal unfolding and aggregation behavior of the protein to allow the ATLAS screen to be optimally designed. |
| تدمد: | 1557-8127 1540-658X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3c585bb5acb0d990ed60e17ee0dab5dc https://doi.org/10.1089/adt.2007.100 |
| رقم الأكسشن: | edsair.doi.dedup.....3c585bb5acb0d990ed60e17ee0dab5dc |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15578127 1540658X |
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