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Fragment screening of cyclin G-associated kinase by weak affinity chromatography
Fragment-based drug discovery (FBDD) has become a new strategy for drug discovery where lead compounds are evolved from small molecules. These fragments form low affinity interactions (dissociation constant (K(D)) = mM − μM) with protein targets, which require fragment screening methods of sufficien...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer-Verlag
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3475971/ https://www.ncbi.nlm.nih.gov/pubmed/22918538 http://dx.doi.org/10.1007/s00216-012-6335-6 |
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author | Meiby, Elinor Knapp, Stefan Elkins, Jonathan M. Ohlson, Sten |
author_facet | Meiby, Elinor Knapp, Stefan Elkins, Jonathan M. Ohlson, Sten |
author_sort | Meiby, Elinor |
collection | PubMed |
description | Fragment-based drug discovery (FBDD) has become a new strategy for drug discovery where lead compounds are evolved from small molecules. These fragments form low affinity interactions (dissociation constant (K(D)) = mM − μM) with protein targets, which require fragment screening methods of sufficient sensitivity. Weak affinity chromatography (WAC) is a promising new technology for fragment screening based on selective retention of fragments by a drug target. Kinases are a major pharmaceutical target, and FBDD has been successfully applied to several of these targets. In this work, we have demonstrated the potential to use WAC in combination with mass spectrometry (MS) detection for fragment screening of a kinase target—cyclin G-associated kinase (GAK). One hundred seventy fragments were selected for WAC screening by virtual screening of a commercial fragment library against the ATP-binding site of five different proteins. GAK protein was immobilized on a capillary HPLC column, and compound binding was characterized by frontal affinity chromatography. Compounds were screened in sets of 13 or 14, in combination with MS detection for enhanced throughput. Seventy-eight fragments (46 %) with K(D) < 200 μM were detected, including a few highly efficient GAK binders (K(D) of 2 μM; ligand efficiency = 0.51). Of special interest is that chiral screening by WAC may be possible, as two stereoisomeric fragments, which both contained one chiral center, demonstrated twin peaks. This ability, in combination with the robustness, sensitivity, and simplicity of WAC makes it a new method for fragment screening of considerable potential. |
format | Online Article Text |
id | pubmed-3475971 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Springer-Verlag |
record_format | MEDLINE/PubMed |
spelling | pubmed-34759712012-10-19 Fragment screening of cyclin G-associated kinase by weak affinity chromatography Meiby, Elinor Knapp, Stefan Elkins, Jonathan M. Ohlson, Sten Anal Bioanal Chem Original Paper Fragment-based drug discovery (FBDD) has become a new strategy for drug discovery where lead compounds are evolved from small molecules. These fragments form low affinity interactions (dissociation constant (K(D)) = mM − μM) with protein targets, which require fragment screening methods of sufficient sensitivity. Weak affinity chromatography (WAC) is a promising new technology for fragment screening based on selective retention of fragments by a drug target. Kinases are a major pharmaceutical target, and FBDD has been successfully applied to several of these targets. In this work, we have demonstrated the potential to use WAC in combination with mass spectrometry (MS) detection for fragment screening of a kinase target—cyclin G-associated kinase (GAK). One hundred seventy fragments were selected for WAC screening by virtual screening of a commercial fragment library against the ATP-binding site of five different proteins. GAK protein was immobilized on a capillary HPLC column, and compound binding was characterized by frontal affinity chromatography. Compounds were screened in sets of 13 or 14, in combination with MS detection for enhanced throughput. Seventy-eight fragments (46 %) with K(D) < 200 μM were detected, including a few highly efficient GAK binders (K(D) of 2 μM; ligand efficiency = 0.51). Of special interest is that chiral screening by WAC may be possible, as two stereoisomeric fragments, which both contained one chiral center, demonstrated twin peaks. This ability, in combination with the robustness, sensitivity, and simplicity of WAC makes it a new method for fragment screening of considerable potential. Springer-Verlag 2012-08-24 2012-11 /pmc/articles/PMC3475971/ /pubmed/22918538 http://dx.doi.org/10.1007/s00216-012-6335-6 Text en © Springer-Verlag 2012 |
spellingShingle | Original Paper Meiby, Elinor Knapp, Stefan Elkins, Jonathan M. Ohlson, Sten Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title | Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title_full | Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title_fullStr | Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title_full_unstemmed | Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title_short | Fragment screening of cyclin G-associated kinase by weak affinity chromatography |
title_sort | fragment screening of cyclin g-associated kinase by weak affinity chromatography |
topic | Original Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3475971/ https://www.ncbi.nlm.nih.gov/pubmed/22918538 http://dx.doi.org/10.1007/s00216-012-6335-6 |
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