Global Kinase Screening. Applications of Frontal Affinity Chromatography Coupled to Mass Spectrometry in Drug Discovery
Utilizing frontal affinity chromatography with mass spectrometry detection (FAC-MS), we have identified novel applications in the discovery of small-molecule hits to protein targets that are difficult if not impossible to accomplish using traditional assays. We demonstrate for the first time an abil...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2005-03, Vol.77 (5), p.1268-1274 |
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| creator | Slon-Usakiewicz, Jacek J Dai, Jin-Rui Ng, William Foster, J. Estelle Deretey, Eugen Toledo-Sherman, Leticia Redden, Peter R Pasternak, Andrew Reid, Neil |
| description | Utilizing frontal affinity chromatography with mass spectrometry detection (FAC-MS), we have identified novel applications in the discovery of small-molecule hits to protein targets that are difficult if not impossible to accomplish using traditional assays. We demonstrate for the first time an ability to distinguish between competitive ligands for the ATP and substrate sites of protein kinase C independently in the same experiment and show that ATP competitive ligands using a functionally inactive receptor tyrosine kinase can be identified. This ability of FAC-MS to simultaneously monitor binding at the ATP and substrate binding sites, as well as measure ligand binding to both active and inactive kinases, suggests that FAC-MS can be used as a “global kinase binding assay”. |
| doi_str_mv | 10.1021/ac048716q |
| format | Article |
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Estelle</creatorcontrib><creatorcontrib>Deretey, Eugen</creatorcontrib><creatorcontrib>Toledo-Sherman, Leticia</creatorcontrib><creatorcontrib>Redden, Peter R</creatorcontrib><creatorcontrib>Pasternak, Andrew</creatorcontrib><creatorcontrib>Reid, Neil</creatorcontrib><title>Global Kinase Screening. Applications of Frontal Affinity Chromatography Coupled to Mass Spectrometry in Drug Discovery</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>Utilizing frontal affinity chromatography with mass spectrometry detection (FAC-MS), we have identified novel applications in the discovery of small-molecule hits to protein targets that are difficult if not impossible to accomplish using traditional assays. We demonstrate for the first time an ability to distinguish between competitive ligands for the ATP and substrate sites of protein kinase C independently in the same experiment and show that ATP competitive ligands using a functionally inactive receptor tyrosine kinase can be identified. This ability of FAC-MS to simultaneously monitor binding at the ATP and substrate binding sites, as well as measure ligand binding to both active and inactive kinases, suggests that FAC-MS can be used as a “global kinase binding assay”.</description><subject>Alkaloids - chemistry</subject><subject>Alkaloids - metabolism</subject><subject>Analytical chemistry</subject><subject>Animals</subject><subject>Benzophenanthridines - chemistry</subject><subject>Benzophenanthridines - metabolism</subject><subject>Binding Sites</subject><subject>Binding, Competitive</subject><subject>Catalytic Domain</subject><subject>Chemistry</subject><subject>Chromatographic methods and physical methods associated with chromatography</subject><subject>Chromatography</subject><subject>Chromatography, Affinity - methods</subject><subject>Drug Evaluation, Preclinical - methods</subject><subject>Drugs</subject><subject>Enzymes</subject><subject>Exact sciences and technology</subject><subject>Humans</subject><subject>Imidazoles - chemistry</subject><subject>Imidazoles - metabolism</subject><subject>Mass spectrometry</subject><subject>Mass Spectrometry - methods</subject><subject>Mice</subject><subject>Molecular Structure</subject><subject>Molecules</subject><subject>Other chromatographic methods</subject><subject>Peptides - chemistry</subject><subject>Peptides - metabolism</subject><subject>Phosphorylation</subject><subject>Protein Binding</subject><subject>Protein Kinase C-alpha - antagonists & inhibitors</subject><subject>Protein Kinase C-alpha - chemistry</subject><subject>Protein Kinase C-alpha - metabolism</subject><subject>Protein Kinase Inhibitors - chemistry</subject><subject>Protein Kinase Inhibitors - metabolism</subject><subject>Protein Kinase Inhibitors - pharmacology</subject><subject>Protein Kinases - chemistry</subject><subject>Protein Kinases - metabolism</subject><subject>Pyridines - chemistry</subject><subject>Pyridines - metabolism</subject><subject>Quinazolines - chemistry</subject><subject>Quinazolines - metabolism</subject><subject>Receptor, EphB2 - antagonists & inhibitors</subject><subject>Receptor, EphB2 - chemistry</subject><subject>Receptor, EphB2 - metabolism</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - metabolism</subject><subject>Spectrometric and optical methods</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2005</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpl0V1v0zAUBmALgVgpXPAHkIU0JC4yjpP4o5dVRwfa-JBatEvLcezOI7UzO2Hrv8dTq1WCK9vyo-P3-CD0lsAZgZJ8UhpqwQm7e4YmhJZQMCHK52gCAFVRcoAT9CqlWwBCgLCX6IRQXpUzYBN0f9GFRnX40nmVDF7paIx3fnOG533fOa0GF3zCweJlDH7Icm6t827Y4cVNDFs1hE1U_U0-hrHvTIuHgL-plPCqN3rIwgxxh53H53Hc4HOXdPhj4u41emFVl8ybwzpFv5af14svxdWPi6-L-VWhahBDURJjgTfczrSFGmrCKAhdVzNj2oYS1lb8cc8bXStbUWY4E0BFKzQFXgqopujDvm4fw91o0iC3OYLpOuVNGJNkvGYlobMM3_8Db8MYfc4mS8IFr1h-aoo-7pGOIaVorOyj26q4kwTk4yjk0yiyfXcoODZb0x7l4e8zOD0AlbTqbFReu3R0jAomCM2u2DuXBvPwdK_i75y-4lSuf67k8vo7gzW_lNfHukqnYxP_B_wLfS2ruw</recordid><startdate>20050301</startdate><enddate>20050301</enddate><creator>Slon-Usakiewicz, Jacek J</creator><creator>Dai, Jin-Rui</creator><creator>Ng, William</creator><creator>Foster, J. 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Applications of Frontal Affinity Chromatography Coupled to Mass Spectrometry in Drug Discovery</title><author>Slon-Usakiewicz, Jacek J ; Dai, Jin-Rui ; Ng, William ; Foster, J. 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Estelle</au><au>Deretey, Eugen</au><au>Toledo-Sherman, Leticia</au><au>Redden, Peter R</au><au>Pasternak, Andrew</au><au>Reid, Neil</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Global Kinase Screening. Applications of Frontal Affinity Chromatography Coupled to Mass Spectrometry in Drug Discovery</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2005-03-01</date><risdate>2005</risdate><volume>77</volume><issue>5</issue><spage>1268</spage><epage>1274</epage><pages>1268-1274</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Utilizing frontal affinity chromatography with mass spectrometry detection (FAC-MS), we have identified novel applications in the discovery of small-molecule hits to protein targets that are difficult if not impossible to accomplish using traditional assays. We demonstrate for the first time an ability to distinguish between competitive ligands for the ATP and substrate sites of protein kinase C independently in the same experiment and show that ATP competitive ligands using a functionally inactive receptor tyrosine kinase can be identified. This ability of FAC-MS to simultaneously monitor binding at the ATP and substrate binding sites, as well as measure ligand binding to both active and inactive kinases, suggests that FAC-MS can be used as a “global kinase binding assay”.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>15732906</pmid><doi>10.1021/ac048716q</doi><tpages>7</tpages></addata></record> |
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| ispartof | Analytical chemistry (Washington), 2005-03, Vol.77 (5), p.1268-1274 |
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| language | eng |
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| subjects | Alkaloids - chemistry Alkaloids - metabolism Analytical chemistry Animals Benzophenanthridines - chemistry Benzophenanthridines - metabolism Binding Sites Binding, Competitive Catalytic Domain Chemistry Chromatographic methods and physical methods associated with chromatography Chromatography Chromatography, Affinity - methods Drug Evaluation, Preclinical - methods Drugs Enzymes Exact sciences and technology Humans Imidazoles - chemistry Imidazoles - metabolism Mass spectrometry Mass Spectrometry - methods Mice Molecular Structure Molecules Other chromatographic methods Peptides - chemistry Peptides - metabolism Phosphorylation Protein Binding Protein Kinase C-alpha - antagonists & inhibitors Protein Kinase C-alpha - chemistry Protein Kinase C-alpha - metabolism Protein Kinase Inhibitors - chemistry Protein Kinase Inhibitors - metabolism Protein Kinase Inhibitors - pharmacology Protein Kinases - chemistry Protein Kinases - metabolism Pyridines - chemistry Pyridines - metabolism Quinazolines - chemistry Quinazolines - metabolism Receptor, EphB2 - antagonists & inhibitors Receptor, EphB2 - chemistry Receptor, EphB2 - metabolism Recombinant Proteins - chemistry Recombinant Proteins - metabolism Spectrometric and optical methods |
| title | Global Kinase Screening. Applications of Frontal Affinity Chromatography Coupled to Mass Spectrometry in Drug Discovery |
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