In Vivo Synthesized Proteins with Monoexponential Fluorescence Decay Kinetics
Tryptophan, when in a protein, typically shows multiexponential fluorescence decay kinetics. Complex kinetics prevents a straightforward interpretation of time-resolved fluorescence protein data, particularly in anisotropy studies or if the effect of a dynamic quencher or a resonance energy transfer...
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| Veröffentlicht in: | Journal of the American Chemical Society 2004-01, Vol.126 (1), p.22-23 |
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| creator | Broos, Jaap Maddalena, Francesco Hesp, Ben H |
| description | Tryptophan, when in a protein, typically shows multiexponential fluorescence decay kinetics. Complex kinetics prevents a straightforward interpretation of time-resolved fluorescence protein data, particularly in anisotropy studies or if the effect of a dynamic quencher or a resonance energy transfer (RET) acceptor is investigated. Here, time-resolved fluorescence data are presented of an isosteric tryptophan analogue, 5-fluorotryptophan, which when biosynthetically incorporated in proteins shows monoexponential decay kinetics. Data are presented indicating that the presence of a fluoro atom at the 5-position suppresses the electron transfer rate from the excited indole moiety to the peptide bond. This process has been related to the multiexponential fluorescence decay of tryptophan in proteins. The monoexponential decay of 5-fluorotryptophan makes it possible to measure simultaneously multiple distances between 5-fluorotryptophan and a RET acceptor. We demonstrate that for an oligomeric protein, consisting of two single-tryptophan-containing subunits, the individual distances between 5-fluorotryptophan and the single substrate binding site can be resolved using a substrate harboring a RET acceptor. |
| doi_str_mv | 10.1021/ja0385585 |
| format | Article |
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Complex kinetics prevents a straightforward interpretation of time-resolved fluorescence protein data, particularly in anisotropy studies or if the effect of a dynamic quencher or a resonance energy transfer (RET) acceptor is investigated. Here, time-resolved fluorescence data are presented of an isosteric tryptophan analogue, 5-fluorotryptophan, which when biosynthetically incorporated in proteins shows monoexponential decay kinetics. Data are presented indicating that the presence of a fluoro atom at the 5-position suppresses the electron transfer rate from the excited indole moiety to the peptide bond. This process has been related to the multiexponential fluorescence decay of tryptophan in proteins. The monoexponential decay of 5-fluorotryptophan makes it possible to measure simultaneously multiple distances between 5-fluorotryptophan and a RET acceptor. 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Psychology ; General aspects, investigation methods ; Kinetics ; Monosaccharide Transport Proteins ; Mutation ; Phosphoenolpyruvate Sugar Phosphotransferase System - chemistry ; Phosphoenolpyruvate Sugar Phosphotransferase System - genetics ; Phosphoenolpyruvate Sugar Phosphotransferase System - metabolism ; Proteins ; Proteins - chemical synthesis ; Proteins - chemistry ; Tryptophan - analogs & derivatives ; Tryptophan - chemistry</subject><ispartof>Journal of the American Chemical Society, 2004-01, Vol.126 (1), p.22-23</ispartof><rights>Copyright © 2004 American Chemical Society</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a451t-5ab0e8456950763e5f1b3c30e922b013e2c23a0aa7d72841eea57f65da3f6e953</citedby><cites>FETCH-LOGICAL-a451t-5ab0e8456950763e5f1b3c30e922b013e2c23a0aa7d72841eea57f65da3f6e953</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/ja0385585$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/ja0385585$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,777,781,2752,27057,27905,27906,56719,56769</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15411426$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14709040$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Broos, Jaap</creatorcontrib><creatorcontrib>Maddalena, Francesco</creatorcontrib><creatorcontrib>Hesp, Ben H</creatorcontrib><title>In Vivo Synthesized Proteins with Monoexponential Fluorescence Decay Kinetics</title><title>Journal of the American Chemical Society</title><addtitle>J. Am. Chem. Soc</addtitle><description>Tryptophan, when in a protein, typically shows multiexponential fluorescence decay kinetics. Complex kinetics prevents a straightforward interpretation of time-resolved fluorescence protein data, particularly in anisotropy studies or if the effect of a dynamic quencher or a resonance energy transfer (RET) acceptor is investigated. Here, time-resolved fluorescence data are presented of an isosteric tryptophan analogue, 5-fluorotryptophan, which when biosynthetically incorporated in proteins shows monoexponential decay kinetics. Data are presented indicating that the presence of a fluoro atom at the 5-position suppresses the electron transfer rate from the excited indole moiety to the peptide bond. This process has been related to the multiexponential fluorescence decay of tryptophan in proteins. The monoexponential decay of 5-fluorotryptophan makes it possible to measure simultaneously multiple distances between 5-fluorotryptophan and a RET acceptor. We demonstrate that for an oligomeric protein, consisting of two single-tryptophan-containing subunits, the individual distances between 5-fluorotryptophan and the single substrate binding site can be resolved using a substrate harboring a RET acceptor.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Escherichia coli - enzymology</subject><subject>Escherichia coli Proteins</subject><subject>Fluorescence</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>General aspects, investigation methods</subject><subject>Kinetics</subject><subject>Monosaccharide Transport Proteins</subject><subject>Mutation</subject><subject>Phosphoenolpyruvate Sugar Phosphotransferase System - chemistry</subject><subject>Phosphoenolpyruvate Sugar Phosphotransferase System - genetics</subject><subject>Phosphoenolpyruvate Sugar Phosphotransferase System - metabolism</subject><subject>Proteins</subject><subject>Proteins - chemical synthesis</subject><subject>Proteins - chemistry</subject><subject>Tryptophan - analogs & derivatives</subject><subject>Tryptophan - chemistry</subject><issn>0002-7863</issn><issn>1520-5126</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkMFOGzEQhq2KqglpD7wA2gtIHLaM7fXu5lglJaCSNhK0ys2aOLPC6cYO9m4hPH0XJSIXTqPRfPpn5mPshMNXDoJfrhBkqVSpPrA-VwJSxUV-xPoAINKizGWPHce46tpMlPwT6_GsgCFk0GfTG5f8sf98crd1zQNF-0LLZBZ8Q9bF5Mk2D8nUO0_PG-_INRbr5KpufaBoyBlKxmRwm_ywjhpr4mf2scI60pd9HbDfV9_vR9fp7a_JzejbbYqZ4k2qcAFUZiofKihySariC2kk0FCIBXBJwgiJgFgsC1FmnAhVUeVqibLKaajkgJ3vcjfBP7YUG7223UF1jY58G3UJUAroHh-wix1ogo8xUKU3wa4xbDUH_epOv7nr2NN9aLtY0_JA7mV1wNkewGiwrgI6Y-OBUxnnmXhdmu44Gxt6fptj-KvzQhZK38_u9OR6LubTn2M9O-SiiXrl2-A6d-8c-B9W-JC_</recordid><startdate>20040114</startdate><enddate>20040114</enddate><creator>Broos, Jaap</creator><creator>Maddalena, Francesco</creator><creator>Hesp, Ben H</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>7X8</scope></search><sort><creationdate>20040114</creationdate><title>In Vivo Synthesized Proteins with Monoexponential Fluorescence Decay Kinetics</title><author>Broos, Jaap ; Maddalena, Francesco ; Hesp, Ben H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a451t-5ab0e8456950763e5f1b3c30e922b013e2c23a0aa7d72841eea57f65da3f6e953</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Escherichia coli - enzymology</topic><topic>Escherichia coli Proteins</topic><topic>Fluorescence</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>General aspects, investigation methods</topic><topic>Kinetics</topic><topic>Monosaccharide Transport Proteins</topic><topic>Mutation</topic><topic>Phosphoenolpyruvate Sugar Phosphotransferase System - chemistry</topic><topic>Phosphoenolpyruvate Sugar Phosphotransferase System - genetics</topic><topic>Phosphoenolpyruvate Sugar Phosphotransferase System - metabolism</topic><topic>Proteins</topic><topic>Proteins - chemical synthesis</topic><topic>Proteins - chemistry</topic><topic>Tryptophan - analogs & derivatives</topic><topic>Tryptophan - chemistry</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Broos, Jaap</creatorcontrib><creatorcontrib>Maddalena, Francesco</creatorcontrib><creatorcontrib>Hesp, Ben H</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>MEDLINE - Academic</collection><jtitle>Journal of the American Chemical Society</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Broos, Jaap</au><au>Maddalena, Francesco</au><au>Hesp, Ben H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In Vivo Synthesized Proteins with Monoexponential Fluorescence Decay Kinetics</atitle><jtitle>Journal of the American Chemical Society</jtitle><addtitle>J. Am. Chem. Soc</addtitle><date>2004-01-14</date><risdate>2004</risdate><volume>126</volume><issue>1</issue><spage>22</spage><epage>23</epage><pages>22-23</pages><issn>0002-7863</issn><eissn>1520-5126</eissn><coden>JACSAT</coden><abstract>Tryptophan, when in a protein, typically shows multiexponential fluorescence decay kinetics. Complex kinetics prevents a straightforward interpretation of time-resolved fluorescence protein data, particularly in anisotropy studies or if the effect of a dynamic quencher or a resonance energy transfer (RET) acceptor is investigated. Here, time-resolved fluorescence data are presented of an isosteric tryptophan analogue, 5-fluorotryptophan, which when biosynthetically incorporated in proteins shows monoexponential decay kinetics. Data are presented indicating that the presence of a fluoro atom at the 5-position suppresses the electron transfer rate from the excited indole moiety to the peptide bond. This process has been related to the multiexponential fluorescence decay of tryptophan in proteins. The monoexponential decay of 5-fluorotryptophan makes it possible to measure simultaneously multiple distances between 5-fluorotryptophan and a RET acceptor. We demonstrate that for an oligomeric protein, consisting of two single-tryptophan-containing subunits, the individual distances between 5-fluorotryptophan and the single substrate binding site can be resolved using a substrate harboring a RET acceptor.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>14709040</pmid><doi>10.1021/ja0385585</doi><tpages>2</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Analytical, structural and metabolic biochemistry Biological and medical sciences Escherichia coli - enzymology Escherichia coli Proteins Fluorescence Fundamental and applied biological sciences. Psychology General aspects, investigation methods Kinetics Monosaccharide Transport Proteins Mutation Phosphoenolpyruvate Sugar Phosphotransferase System - chemistry Phosphoenolpyruvate Sugar Phosphotransferase System - genetics Phosphoenolpyruvate Sugar Phosphotransferase System - metabolism Proteins Proteins - chemical synthesis Proteins - chemistry Tryptophan - analogs & derivatives Tryptophan - chemistry |
| title | In Vivo Synthesized Proteins with Monoexponential Fluorescence Decay Kinetics |
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