A Molecular Docking Strategy Identifies Eosin B as a Non-active Site Inhibitor of Protozoal Bifunctional Thymidylate Synthase-Dihydrofolate Reductase
Protozoal parasites are unusual in that their thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes exist on a single polypeptide. In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS...
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| Veröffentlicht in: | The Journal of biological chemistry 2003-04, Vol.278 (16), p.14092-14100 |
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| container_title | The Journal of biological chemistry |
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| creator | Atreya, Chloé E. Johnson, Eric F. Irwin, John J. Dow, Antonia Massimine, Kristen M. Coppens, Isabelle Stempliuk, Valeska Beverley, Stephen Joiner, Keith A. Shoichet, Brian K. Anderson, Karen S. |
| description | Protozoal parasites are unusual in that their thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes exist on a single polypeptide. In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS-DHFR, we used molecular docking to screen for inhibitors targeting the shallow groove connecting the two active sites. Eosin B is a 100 μm non-active site inhibitor of Leishmania major TS-DHFR identified by molecular docking. Eosin B slows both the TS and DHFR reaction rates. When Arg-283, a key residue to which eosin B is predicted to bind, is mutated to glutamate, however, eosin B only minimally inhibits the TS-DHFR reaction. Additionally, eosin B was found to be a 180 μm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays. |
| doi_str_mv | 10.1074/jbc.M212690200 |
| format | Article |
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In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS-DHFR, we used molecular docking to screen for inhibitors targeting the shallow groove connecting the two active sites. Eosin B is a 100 μm non-active site inhibitor of Leishmania major TS-DHFR identified by molecular docking. Eosin B slows both the TS and DHFR reaction rates. When Arg-283, a key residue to which eosin B is predicted to bind, is mutated to glutamate, however, eosin B only minimally inhibits the TS-DHFR reaction. Additionally, eosin B was found to be a 180 μm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M212690200</identifier><identifier>PMID: 12556445</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Antiprotozoal Agents - pharmacology ; Arginine - chemistry ; Binding Sites ; CHO Cells ; Chromatography, High Pressure Liquid ; Cricetinae ; Dose-Response Relationship, Drug ; Enzyme Inhibitors - pharmacology ; Eosine I Bluish ; Fluoresceins - chemistry ; Fluoresceins - metabolism ; Fluoresceins - pharmacology ; Glutamic Acid - chemistry ; Hydrogen-Ion Concentration ; Inhibitory Concentration 50 ; Kinetics ; Leishmania major - metabolism ; Models, Molecular ; Multienzyme Complexes - chemistry ; Multienzyme Complexes - metabolism ; Protein Binding ; Protein Structure, Tertiary ; Tetrahydrofolate Dehydrogenase - chemistry ; Tetrahydrofolate Dehydrogenase - metabolism ; Thymidylate Synthase - chemistry ; Thymidylate Synthase - metabolism ; Time Factors ; Toxoplasma - metabolism</subject><ispartof>The Journal of biological chemistry, 2003-04, Vol.278 (16), p.14092-14100</ispartof><rights>2003 © 2003 ASBMB. 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Additionally, eosin B was found to be a 180 μm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays.</description><subject>Animals</subject><subject>Antiprotozoal Agents - pharmacology</subject><subject>Arginine - chemistry</subject><subject>Binding Sites</subject><subject>CHO Cells</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Cricetinae</subject><subject>Dose-Response Relationship, Drug</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Eosine I Bluish</subject><subject>Fluoresceins - chemistry</subject><subject>Fluoresceins - metabolism</subject><subject>Fluoresceins - pharmacology</subject><subject>Glutamic Acid - chemistry</subject><subject>Hydrogen-Ion Concentration</subject><subject>Inhibitory Concentration 50</subject><subject>Kinetics</subject><subject>Leishmania major - metabolism</subject><subject>Models, Molecular</subject><subject>Multienzyme Complexes - chemistry</subject><subject>Multienzyme Complexes - metabolism</subject><subject>Protein Binding</subject><subject>Protein Structure, Tertiary</subject><subject>Tetrahydrofolate Dehydrogenase - chemistry</subject><subject>Tetrahydrofolate Dehydrogenase - metabolism</subject><subject>Thymidylate Synthase - chemistry</subject><subject>Thymidylate Synthase - metabolism</subject><subject>Time Factors</subject><subject>Toxoplasma - metabolism</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kU1v1DAQhi0EokvhyhH5gLhlsR3n69gvykotILZI3CzHHm9cErvYTqvwP_i_GHalnpjLaF4982o0L0KvKVlT0vD3t71aXzPK6o4wQp6gFSVtWZQV_f4UrQhhtOhY1R6hFzHekly8o8_REWVVVXNerdDvE3ztR1DzKAM-9-qHdTu8TUEm2C14o8ElayxEfOGjdfgUy4gl_uRdIVWy94C3NgHeuMH2NvmAvcFfgk_-l5cjPrVmdhnzLg83wzJZvYzZGW8XlwYZoTi3w6KDN_6f_BX0rFLWX6JnRo4RXh36Mfr24eLm7GNx9flyc3ZyVSjOSSr6Hgi0HeFtT3uqmSFEMw512fISjKbKlKaSWkJtZMWU6RRrtKEVg141pFblMXq3970L_ucMMYnJRgXjKB34OQraNrSpmjaD6z2ogo8xgBF3wU4yLIIS8TcIkYMQj0HkhTcH57mfQD_ih89n4O0eGOxueLABRG-9GmASrGkFrQXlpGMZa_cY5DfcWwgiKgtOgc4rKgnt7f9O-AMK96Yg</recordid><startdate>20030418</startdate><enddate>20030418</enddate><creator>Atreya, Chloé E.</creator><creator>Johnson, Eric F.</creator><creator>Irwin, John J.</creator><creator>Dow, Antonia</creator><creator>Massimine, Kristen M.</creator><creator>Coppens, Isabelle</creator><creator>Stempliuk, Valeska</creator><creator>Beverley, Stephen</creator><creator>Joiner, Keith A.</creator><creator>Shoichet, Brian K.</creator><creator>Anderson, Karen S.</creator><general>Elsevier Inc</general><general>American Society for Biochemistry and Molecular Biology</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>M7N</scope></search><sort><creationdate>20030418</creationdate><title>A Molecular Docking Strategy Identifies Eosin B as a Non-active Site Inhibitor of Protozoal Bifunctional Thymidylate Synthase-Dihydrofolate Reductase</title><author>Atreya, Chloé E. ; Johnson, Eric F. ; Irwin, John J. ; Dow, Antonia ; Massimine, Kristen M. ; Coppens, Isabelle ; Stempliuk, Valeska ; Beverley, Stephen ; Joiner, Keith A. ; Shoichet, Brian K. ; Anderson, Karen S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-bbe0e89048b1b1d2f00d24e63843efd1cf3f5adae6fa52cf9c27df152ebc706c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>Antiprotozoal Agents - pharmacology</topic><topic>Arginine - chemistry</topic><topic>Binding Sites</topic><topic>CHO Cells</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Cricetinae</topic><topic>Dose-Response Relationship, Drug</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Eosine I Bluish</topic><topic>Fluoresceins - chemistry</topic><topic>Fluoresceins - metabolism</topic><topic>Fluoresceins - pharmacology</topic><topic>Glutamic Acid - chemistry</topic><topic>Hydrogen-Ion Concentration</topic><topic>Inhibitory Concentration 50</topic><topic>Kinetics</topic><topic>Leishmania major - metabolism</topic><topic>Models, Molecular</topic><topic>Multienzyme Complexes - chemistry</topic><topic>Multienzyme Complexes - metabolism</topic><topic>Protein Binding</topic><topic>Protein Structure, Tertiary</topic><topic>Tetrahydrofolate Dehydrogenase - chemistry</topic><topic>Tetrahydrofolate Dehydrogenase - metabolism</topic><topic>Thymidylate Synthase - chemistry</topic><topic>Thymidylate Synthase - metabolism</topic><topic>Time Factors</topic><topic>Toxoplasma - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Atreya, Chloé E.</creatorcontrib><creatorcontrib>Johnson, Eric F.</creatorcontrib><creatorcontrib>Irwin, John J.</creatorcontrib><creatorcontrib>Dow, Antonia</creatorcontrib><creatorcontrib>Massimine, Kristen M.</creatorcontrib><creatorcontrib>Coppens, Isabelle</creatorcontrib><creatorcontrib>Stempliuk, Valeska</creatorcontrib><creatorcontrib>Beverley, Stephen</creatorcontrib><creatorcontrib>Joiner, Keith A.</creatorcontrib><creatorcontrib>Shoichet, Brian K.</creatorcontrib><creatorcontrib>Anderson, Karen S.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Atreya, Chloé E.</au><au>Johnson, Eric F.</au><au>Irwin, John J.</au><au>Dow, Antonia</au><au>Massimine, Kristen M.</au><au>Coppens, Isabelle</au><au>Stempliuk, Valeska</au><au>Beverley, Stephen</au><au>Joiner, Keith A.</au><au>Shoichet, Brian K.</au><au>Anderson, Karen S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A Molecular Docking Strategy Identifies Eosin B as a Non-active Site Inhibitor of Protozoal Bifunctional Thymidylate Synthase-Dihydrofolate Reductase</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>2003-04-18</date><risdate>2003</risdate><volume>278</volume><issue>16</issue><spage>14092</spage><epage>14100</epage><pages>14092-14100</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>Protozoal parasites are unusual in that their thymidylate synthase (TS) and dihydrofolate reductase (DHFR) enzymes exist on a single polypeptide. In an effort to probe the possibility of substrate channeling between the TS and DHFR active sites and to identify inhibitors specific for bifunctional TS-DHFR, we used molecular docking to screen for inhibitors targeting the shallow groove connecting the two active sites. Eosin B is a 100 μm non-active site inhibitor of Leishmania major TS-DHFR identified by molecular docking. Eosin B slows both the TS and DHFR reaction rates. When Arg-283, a key residue to which eosin B is predicted to bind, is mutated to glutamate, however, eosin B only minimally inhibits the TS-DHFR reaction. Additionally, eosin B was found to be a 180 μm inhibitor of Toxoplasma gondii in both biochemical and cell culture assays.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>12556445</pmid><doi>10.1074/jbc.M212690200</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The Journal of biological chemistry, 2003-04, Vol.278 (16), p.14092-14100 |
| issn | 0021-9258 1083-351X |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animals Antiprotozoal Agents - pharmacology Arginine - chemistry Binding Sites CHO Cells Chromatography, High Pressure Liquid Cricetinae Dose-Response Relationship, Drug Enzyme Inhibitors - pharmacology Eosine I Bluish Fluoresceins - chemistry Fluoresceins - metabolism Fluoresceins - pharmacology Glutamic Acid - chemistry Hydrogen-Ion Concentration Inhibitory Concentration 50 Kinetics Leishmania major - metabolism Models, Molecular Multienzyme Complexes - chemistry Multienzyme Complexes - metabolism Protein Binding Protein Structure, Tertiary Tetrahydrofolate Dehydrogenase - chemistry Tetrahydrofolate Dehydrogenase - metabolism Thymidylate Synthase - chemistry Thymidylate Synthase - metabolism Time Factors Toxoplasma - metabolism |
| title | A Molecular Docking Strategy Identifies Eosin B as a Non-active Site Inhibitor of Protozoal Bifunctional Thymidylate Synthase-Dihydrofolate Reductase |
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