Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library
A simple procedure for selection of tight-binding inhibitors of mutant dihydrofolate reductases from Plasmodium falciparum (PfDHFRs) based on preferential binding to the enzyme immobilized on a Sepharose column has been described. PfDHFRs with a cysteine residue at the C-terminal have been prepared...
Gespeichert in:
| Veröffentlicht in: | Analytical chemistry (Washington) 2007-07, Vol.79 (13), p.5006-5012 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 5012 |
|---|---|
| container_issue | 13 |
| container_start_page | 5006 |
| container_title | Analytical chemistry (Washington) |
| container_volume | 79 |
| creator | Thongpanchang, Chawanee Taweechai, Supannee Kamchonwongpaisan, Sumalee Yuthavong, Yongyuth Thebtaranonth, Yodhathai |
| description | A simple procedure for selection of tight-binding inhibitors of mutant dihydrofolate reductases from Plasmodium falciparum (PfDHFRs) based on preferential binding to the enzyme immobilized on a Sepharose column has been described. PfDHFRs with a cysteine residue at the C-terminal have been prepared in order to immobilize to a thiopropyl-Sepharose gel via S−S linkage. The amount of immobilized DHFRs was estimated to be 4−5 mg/g of dried gel, and the activities of bound DHFRs were comparable to that of free enzymes. The prepared immobilized enzyme has been used for the selection of tight-binding inhibitors from combinatorial libraries, based on the affinities of each ligand with the enzyme. Free ligands were then identified and analyzed quantitatively by high-performance liquid chromatography−mass spectrometry, and the components with high binding affinity of the library could thus be realized. Results could be confirmed by quantitative analysis of the bound ligands released from the enzyme by guanidine hydrochloride treatment. |
| doi_str_mv | 10.1021/ac070215s |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_70678568</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1298736841</sourcerecordid><originalsourceid>FETCH-LOGICAL-a439t-54e71f054dd4c6560abc6270c3e55c91a521cc69fcb95a7ac802620b621841e03</originalsourceid><addsrcrecordid>eNqF0c1u1DAUBeAIgei0sOAFkIUEoovAtRP_ZEmnUEYaoGKGDRvrxnE6Lkk82IlEeQ1eGFczdCRYsLJkfzrXRzfLnlB4RYHR12hAppPHe9mMcga5UIrdz2YAUORMAhxlxzFeA1AKVDzMjqjkBcgSZtmvRd_72nXuJ47OD8S35AN2GBx25OVlh7H3jZt60mJn3BbD1J-Sc7e5aYJvfYejJZ9tM5kRoyWtD2TcWLKynTV_0tbuajPmZ25o3HBFFsPG1W70IZI2-J7MfV-7AdPF7cClqwOGm0fZgzQu2sf78yT78u7tev4-X366WMzfLHMsi2rMeWklbYGXTVMawQVgbURqawrLuakockaNEVVr6oqjRKOACQa1YFSV1EJxkr3Y5W6D_z7ZOOreRWO7Dgfrp6glCKm4UP-FtFKVYlIm-OwveO2nMKQSmlGpFJQFT-h0h0zwMQbb6m1wfeqtKejbfeq7fSb7dB841b1tDnK_wASe7wFGg10bcDAuHpyqGFBeJZfvnIuj_XH3juGbFrKQXK8vV3r1cX6xnn9d67NDLpp4KPHvB38D1BbD_g</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>217880435</pqid></control><display><type>article</type><title>Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library</title><source>MEDLINE</source><source>ACS Publications</source><creator>Thongpanchang, Chawanee ; Taweechai, Supannee ; Kamchonwongpaisan, Sumalee ; Yuthavong, Yongyuth ; Thebtaranonth, Yodhathai</creator><creatorcontrib>Thongpanchang, Chawanee ; Taweechai, Supannee ; Kamchonwongpaisan, Sumalee ; Yuthavong, Yongyuth ; Thebtaranonth, Yodhathai</creatorcontrib><description>A simple procedure for selection of tight-binding inhibitors of mutant dihydrofolate reductases from Plasmodium falciparum (PfDHFRs) based on preferential binding to the enzyme immobilized on a Sepharose column has been described. PfDHFRs with a cysteine residue at the C-terminal have been prepared in order to immobilize to a thiopropyl-Sepharose gel via S−S linkage. The amount of immobilized DHFRs was estimated to be 4−5 mg/g of dried gel, and the activities of bound DHFRs were comparable to that of free enzymes. The prepared immobilized enzyme has been used for the selection of tight-binding inhibitors from combinatorial libraries, based on the affinities of each ligand with the enzyme. Free ligands were then identified and analyzed quantitatively by high-performance liquid chromatography−mass spectrometry, and the components with high binding affinity of the library could thus be realized. Results could be confirmed by quantitative analysis of the bound ligands released from the enzyme by guanidine hydrochloride treatment.</description><identifier>ISSN: 0003-2700</identifier><identifier>EISSN: 1520-6882</identifier><identifier>DOI: 10.1021/ac070215s</identifier><identifier>PMID: 17530740</identifier><identifier>CODEN: ANCHAM</identifier><language>eng</language><publisher>Washington, DC: American Chemical Society</publisher><subject>Amino Acid Sequence ; Analytical chemistry ; Animals ; Base Sequence ; Binding Sites ; Biochemistry ; Chemistry ; Chromatographic methods and physical methods associated with chromatography ; Chromatography ; Chromatography, High Pressure Liquid - methods ; Combinatorial Chemistry Techniques ; Cysteine - chemistry ; Enzymes ; Enzymes, Immobilized ; Exact sciences and technology ; Folic Acid Antagonists - isolation & purification ; Guanidine - pharmacology ; Ligands ; Mass spectrometry ; Mass Spectrometry - methods ; Molecular Sequence Data ; Other chromatographic methods ; Parasitic protozoa ; Peptide Library ; Plasmodium falciparum ; Plasmodium falciparum - enzymology ; Plasmodium falciparum - genetics ; Reproducibility of Results ; Sensitivity and Specificity ; Sepharose - chemistry ; Spectrometric and optical methods ; Sulfhydryl Compounds - chemistry ; Sulfides - chemistry ; Tetrahydrofolate Dehydrogenase - chemistry ; Tetrahydrofolate Dehydrogenase - genetics ; Tetrahydrofolate Dehydrogenase - metabolism</subject><ispartof>Analytical chemistry (Washington), 2007-07, Vol.79 (13), p.5006-5012</ispartof><rights>Copyright © 2007 American Chemical Society</rights><rights>2007 INIST-CNRS</rights><rights>Copyright American Chemical Society Jul 1, 2007</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a439t-54e71f054dd4c6560abc6270c3e55c91a521cc69fcb95a7ac802620b621841e03</citedby><cites>FETCH-LOGICAL-a439t-54e71f054dd4c6560abc6270c3e55c91a521cc69fcb95a7ac802620b621841e03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/ac070215s$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ac070215s$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18920159$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17530740$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Thongpanchang, Chawanee</creatorcontrib><creatorcontrib>Taweechai, Supannee</creatorcontrib><creatorcontrib>Kamchonwongpaisan, Sumalee</creatorcontrib><creatorcontrib>Yuthavong, Yongyuth</creatorcontrib><creatorcontrib>Thebtaranonth, Yodhathai</creatorcontrib><title>Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library</title><title>Analytical chemistry (Washington)</title><addtitle>Anal. Chem</addtitle><description>A simple procedure for selection of tight-binding inhibitors of mutant dihydrofolate reductases from Plasmodium falciparum (PfDHFRs) based on preferential binding to the enzyme immobilized on a Sepharose column has been described. PfDHFRs with a cysteine residue at the C-terminal have been prepared in order to immobilize to a thiopropyl-Sepharose gel via S−S linkage. The amount of immobilized DHFRs was estimated to be 4−5 mg/g of dried gel, and the activities of bound DHFRs were comparable to that of free enzymes. The prepared immobilized enzyme has been used for the selection of tight-binding inhibitors from combinatorial libraries, based on the affinities of each ligand with the enzyme. Free ligands were then identified and analyzed quantitatively by high-performance liquid chromatography−mass spectrometry, and the components with high binding affinity of the library could thus be realized. Results could be confirmed by quantitative analysis of the bound ligands released from the enzyme by guanidine hydrochloride treatment.</description><subject>Amino Acid Sequence</subject><subject>Analytical chemistry</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Biochemistry</subject><subject>Chemistry</subject><subject>Chromatographic methods and physical methods associated with chromatography</subject><subject>Chromatography</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Combinatorial Chemistry Techniques</subject><subject>Cysteine - chemistry</subject><subject>Enzymes</subject><subject>Enzymes, Immobilized</subject><subject>Exact sciences and technology</subject><subject>Folic Acid Antagonists - isolation & purification</subject><subject>Guanidine - pharmacology</subject><subject>Ligands</subject><subject>Mass spectrometry</subject><subject>Mass Spectrometry - methods</subject><subject>Molecular Sequence Data</subject><subject>Other chromatographic methods</subject><subject>Parasitic protozoa</subject><subject>Peptide Library</subject><subject>Plasmodium falciparum</subject><subject>Plasmodium falciparum - enzymology</subject><subject>Plasmodium falciparum - genetics</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Sepharose - chemistry</subject><subject>Spectrometric and optical methods</subject><subject>Sulfhydryl Compounds - chemistry</subject><subject>Sulfides - chemistry</subject><subject>Tetrahydrofolate Dehydrogenase - chemistry</subject><subject>Tetrahydrofolate Dehydrogenase - genetics</subject><subject>Tetrahydrofolate Dehydrogenase - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0c1u1DAUBeAIgei0sOAFkIUEoovAtRP_ZEmnUEYaoGKGDRvrxnE6Lkk82IlEeQ1eGFczdCRYsLJkfzrXRzfLnlB4RYHR12hAppPHe9mMcga5UIrdz2YAUORMAhxlxzFeA1AKVDzMjqjkBcgSZtmvRd_72nXuJ47OD8S35AN2GBx25OVlh7H3jZt60mJn3BbD1J-Sc7e5aYJvfYejJZ9tM5kRoyWtD2TcWLKynTV_0tbuajPmZ25o3HBFFsPG1W70IZI2-J7MfV-7AdPF7cClqwOGm0fZgzQu2sf78yT78u7tev4-X366WMzfLHMsi2rMeWklbYGXTVMawQVgbURqawrLuakockaNEVVr6oqjRKOACQa1YFSV1EJxkr3Y5W6D_z7ZOOreRWO7Dgfrp6glCKm4UP-FtFKVYlIm-OwveO2nMKQSmlGpFJQFT-h0h0zwMQbb6m1wfeqtKejbfeq7fSb7dB841b1tDnK_wASe7wFGg10bcDAuHpyqGFBeJZfvnIuj_XH3juGbFrKQXK8vV3r1cX6xnn9d67NDLpp4KPHvB38D1BbD_g</recordid><startdate>20070701</startdate><enddate>20070701</enddate><creator>Thongpanchang, Chawanee</creator><creator>Taweechai, Supannee</creator><creator>Kamchonwongpaisan, Sumalee</creator><creator>Yuthavong, Yongyuth</creator><creator>Thebtaranonth, Yodhathai</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>IQODW</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>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7TA</scope><scope>7TB</scope><scope>7TM</scope><scope>7U5</scope><scope>7U7</scope><scope>7U9</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>H94</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>F1W</scope><scope>H95</scope><scope>H97</scope><scope>L.G</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>20070701</creationdate><title>Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library</title><author>Thongpanchang, Chawanee ; Taweechai, Supannee ; Kamchonwongpaisan, Sumalee ; Yuthavong, Yongyuth ; Thebtaranonth, Yodhathai</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a439t-54e71f054dd4c6560abc6270c3e55c91a521cc69fcb95a7ac802620b621841e03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Amino Acid Sequence</topic><topic>Analytical chemistry</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Biochemistry</topic><topic>Chemistry</topic><topic>Chromatographic methods and physical methods associated with chromatography</topic><topic>Chromatography</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Combinatorial Chemistry Techniques</topic><topic>Cysteine - chemistry</topic><topic>Enzymes</topic><topic>Enzymes, Immobilized</topic><topic>Exact sciences and technology</topic><topic>Folic Acid Antagonists - isolation & purification</topic><topic>Guanidine - pharmacology</topic><topic>Ligands</topic><topic>Mass spectrometry</topic><topic>Mass Spectrometry - methods</topic><topic>Molecular Sequence Data</topic><topic>Other chromatographic methods</topic><topic>Parasitic protozoa</topic><topic>Peptide Library</topic><topic>Plasmodium falciparum</topic><topic>Plasmodium falciparum - enzymology</topic><topic>Plasmodium falciparum - genetics</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Sepharose - chemistry</topic><topic>Spectrometric and optical methods</topic><topic>Sulfhydryl Compounds - chemistry</topic><topic>Sulfides - chemistry</topic><topic>Tetrahydrofolate Dehydrogenase - chemistry</topic><topic>Tetrahydrofolate Dehydrogenase - genetics</topic><topic>Tetrahydrofolate Dehydrogenase - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Thongpanchang, Chawanee</creatorcontrib><creatorcontrib>Taweechai, Supannee</creatorcontrib><creatorcontrib>Kamchonwongpaisan, Sumalee</creatorcontrib><creatorcontrib>Yuthavong, Yongyuth</creatorcontrib><creatorcontrib>Thebtaranonth, Yodhathai</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Thongpanchang, Chawanee</au><au>Taweechai, Supannee</au><au>Kamchonwongpaisan, Sumalee</au><au>Yuthavong, Yongyuth</au><au>Thebtaranonth, Yodhathai</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2007-07-01</date><risdate>2007</risdate><volume>79</volume><issue>13</issue><spage>5006</spage><epage>5012</epage><pages>5006-5012</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>A simple procedure for selection of tight-binding inhibitors of mutant dihydrofolate reductases from Plasmodium falciparum (PfDHFRs) based on preferential binding to the enzyme immobilized on a Sepharose column has been described. PfDHFRs with a cysteine residue at the C-terminal have been prepared in order to immobilize to a thiopropyl-Sepharose gel via S−S linkage. The amount of immobilized DHFRs was estimated to be 4−5 mg/g of dried gel, and the activities of bound DHFRs were comparable to that of free enzymes. The prepared immobilized enzyme has been used for the selection of tight-binding inhibitors from combinatorial libraries, based on the affinities of each ligand with the enzyme. Free ligands were then identified and analyzed quantitatively by high-performance liquid chromatography−mass spectrometry, and the components with high binding affinity of the library could thus be realized. Results could be confirmed by quantitative analysis of the bound ligands released from the enzyme by guanidine hydrochloride treatment.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>17530740</pmid><doi>10.1021/ac070215s</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2700 |
| ispartof | Analytical chemistry (Washington), 2007-07, Vol.79 (13), p.5006-5012 |
| issn | 0003-2700 1520-6882 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_70678568 |
| source | MEDLINE; ACS Publications |
| subjects | Amino Acid Sequence Analytical chemistry Animals Base Sequence Binding Sites Biochemistry Chemistry Chromatographic methods and physical methods associated with chromatography Chromatography Chromatography, High Pressure Liquid - methods Combinatorial Chemistry Techniques Cysteine - chemistry Enzymes Enzymes, Immobilized Exact sciences and technology Folic Acid Antagonists - isolation & purification Guanidine - pharmacology Ligands Mass spectrometry Mass Spectrometry - methods Molecular Sequence Data Other chromatographic methods Parasitic protozoa Peptide Library Plasmodium falciparum Plasmodium falciparum - enzymology Plasmodium falciparum - genetics Reproducibility of Results Sensitivity and Specificity Sepharose - chemistry Spectrometric and optical methods Sulfhydryl Compounds - chemistry Sulfides - chemistry Tetrahydrofolate Dehydrogenase - chemistry Tetrahydrofolate Dehydrogenase - genetics Tetrahydrofolate Dehydrogenase - metabolism |
| title | Immobilization of Malarial (Plasmodium falciparum) Dihydrofolate Reductase for the Selection of Tight-Binding Inhibitors from Combinatorial Library |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-22T16%3A23%3A01IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Immobilization%20of%20Malarial%20(Plasmodium%20falciparum)%20Dihydrofolate%20Reductase%20for%20the%20Selection%20of%20Tight-Binding%20Inhibitors%20from%20Combinatorial%20Library&rft.jtitle=Analytical%20chemistry%20(Washington)&rft.au=Thongpanchang,%20Chawanee&rft.date=2007-07-01&rft.volume=79&rft.issue=13&rft.spage=5006&rft.epage=5012&rft.pages=5006-5012&rft.issn=0003-2700&rft.eissn=1520-6882&rft.coden=ANCHAM&rft_id=info:doi/10.1021/ac070215s&rft_dat=%3Cproquest_cross%3E1298736841%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=217880435&rft_id=info:pmid/17530740&rfr_iscdi=true |