Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys)
Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro...
Gespeichert in:
| Veröffentlicht in: | Analytical biochemistry 2002-08, Vol.307 (2), p.252-257 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 257 |
|---|---|
| container_issue | 2 |
| container_start_page | 252 |
| container_title | Analytical biochemistry |
| container_volume | 307 |
| creator | Lien, Linda Ananda, P Seneviratne, K Jaikaran, Anna S I Andrew Woolley, G |
| description | Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site. |
| doi_str_mv | 10.1016/S0003-2697(02)00035-0 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_proquest_miscellaneous_72040139</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>72040139</sourcerecordid><originalsourceid>FETCH-LOGICAL-p207t-cb3cd65a5a72fa11980a13fed2723cda1e8178057c67eda771322d911b560c903</originalsourceid><addsrcrecordid>eNo9kE1Lw0AQhveg2Fr9CcqepD2szmyabHMsxS8oClbPYbOZ6EqSjdmNkH9vitXT8Mw8DC8vYxcI1wiY3OwAIBIySdUc5GIPsYAjNv1fT9ip958AiMs4OWETlBKkXOKU5TsbSPiWjC2t4bl1fmjCB4URbGNc17pOB-sa7kqueVn1riNvqAk86PdRCY63nQtkG8-_reZm8HsgEV6e1vPN4Bdn7LjUlafzw5yxt7vb182D2D7fP27WW9FKUEGYPDJFEutYK1lqxHQFGqOSCqnkeNFIK1QriJVJFBVaKYykLFLEPE7ApBDN2NXv3zHPV08-ZLUdk1aVbsj1PlMSloBROoqXB7HPayqytrO17obsr5XoBwF_Y7o</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>72040139</pqid></control><display><type>article</type><title>Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys)</title><source>MEDLINE</source><source>Access via ScienceDirect (Elsevier)</source><creator>Lien, Linda ; Ananda, P ; Seneviratne, K ; Jaikaran, Anna S I ; Andrew Woolley, G</creator><creatorcontrib>Lien, Linda ; Ananda, P ; Seneviratne, K ; Jaikaran, Anna S I ; Andrew Woolley, G</creatorcontrib><description>Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site.</description><identifier>ISSN: 0003-2697</identifier><identifier>DOI: 10.1016/S0003-2697(02)00035-0</identifier><identifier>PMID: 12202241</identifier><language>eng</language><publisher>United States</publisher><subject>Base Sequence ; Boron Compounds - metabolism ; Cell-Free System ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Fluorescent Dyes - metabolism ; Models, Molecular ; Molecular Sequence Data ; Nucleic Acid Conformation ; Proteins - chemistry ; Proteins - metabolism ; RNA, Transfer, Cys - genetics ; RNA, Transfer, Cys - metabolism ; Spectrophotometry, Ultraviolet</subject><ispartof>Analytical biochemistry, 2002-08, Vol.307 (2), p.252-257</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12202241$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lien, Linda</creatorcontrib><creatorcontrib>Ananda, P</creatorcontrib><creatorcontrib>Seneviratne, K</creatorcontrib><creatorcontrib>Jaikaran, Anna S I</creatorcontrib><creatorcontrib>Andrew Woolley, G</creatorcontrib><title>Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys)</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site.</description><subject>Base Sequence</subject><subject>Boron Compounds - metabolism</subject><subject>Cell-Free System</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Escherichia coli</subject><subject>Fluorescent Dyes - metabolism</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Nucleic Acid Conformation</subject><subject>Proteins - chemistry</subject><subject>Proteins - metabolism</subject><subject>RNA, Transfer, Cys - genetics</subject><subject>RNA, Transfer, Cys - metabolism</subject><subject>Spectrophotometry, Ultraviolet</subject><issn>0003-2697</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kE1Lw0AQhveg2Fr9CcqepD2szmyabHMsxS8oClbPYbOZ6EqSjdmNkH9vitXT8Mw8DC8vYxcI1wiY3OwAIBIySdUc5GIPsYAjNv1fT9ip958AiMs4OWETlBKkXOKU5TsbSPiWjC2t4bl1fmjCB4URbGNc17pOB-sa7kqueVn1riNvqAk86PdRCY63nQtkG8-_reZm8HsgEV6e1vPN4Bdn7LjUlafzw5yxt7vb182D2D7fP27WW9FKUEGYPDJFEutYK1lqxHQFGqOSCqnkeNFIK1QriJVJFBVaKYykLFLEPE7ApBDN2NXv3zHPV08-ZLUdk1aVbsj1PlMSloBROoqXB7HPayqytrO17obsr5XoBwF_Y7o</recordid><startdate>20020815</startdate><enddate>20020815</enddate><creator>Lien, Linda</creator><creator>Ananda, P</creator><creator>Seneviratne, K</creator><creator>Jaikaran, Anna S I</creator><creator>Andrew Woolley, G</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>20020815</creationdate><title>Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys)</title><author>Lien, Linda ; Ananda, P ; Seneviratne, K ; Jaikaran, Anna S I ; Andrew Woolley, G</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p207t-cb3cd65a5a72fa11980a13fed2723cda1e8178057c67eda771322d911b560c903</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Base Sequence</topic><topic>Boron Compounds - metabolism</topic><topic>Cell-Free System</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Escherichia coli</topic><topic>Fluorescent Dyes - metabolism</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Nucleic Acid Conformation</topic><topic>Proteins - chemistry</topic><topic>Proteins - metabolism</topic><topic>RNA, Transfer, Cys - genetics</topic><topic>RNA, Transfer, Cys - metabolism</topic><topic>Spectrophotometry, Ultraviolet</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lien, Linda</creatorcontrib><creatorcontrib>Ananda, P</creatorcontrib><creatorcontrib>Seneviratne, K</creatorcontrib><creatorcontrib>Jaikaran, Anna S I</creatorcontrib><creatorcontrib>Andrew Woolley, G</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lien, Linda</au><au>Ananda, P</au><au>Seneviratne, K</au><au>Jaikaran, Anna S I</au><au>Andrew Woolley, G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys)</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>2002-08-15</date><risdate>2002</risdate><volume>307</volume><issue>2</issue><spage>252</spage><epage>257</epage><pages>252-257</pages><issn>0003-2697</issn><abstract>Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site.</abstract><cop>United States</cop><pmid>12202241</pmid><doi>10.1016/S0003-2697(02)00035-0</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0003-2697 |
| ispartof | Analytical biochemistry, 2002-08, Vol.307 (2), p.252-257 |
| issn | 0003-2697 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_72040139 |
| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Base Sequence Boron Compounds - metabolism Cell-Free System Electrophoresis, Polyacrylamide Gel Escherichia coli Fluorescent Dyes - metabolism Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Proteins - chemistry Proteins - metabolism RNA, Transfer, Cys - genetics RNA, Transfer, Cys - metabolism Spectrophotometry, Ultraviolet |
| title | Site-specific biosynthetic incorporation of a fluorescent tag into proteins via cysteine-tRNA(Cys) |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-21T07%3A54%3A49IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Site-specific%20biosynthetic%20incorporation%20of%20a%20fluorescent%20tag%20into%20proteins%20via%20cysteine-tRNA(Cys)&rft.jtitle=Analytical%20biochemistry&rft.au=Lien,%20Linda&rft.date=2002-08-15&rft.volume=307&rft.issue=2&rft.spage=252&rft.epage=257&rft.pages=252-257&rft.issn=0003-2697&rft_id=info:doi/10.1016/S0003-2697(02)00035-0&rft_dat=%3Cproquest_pubme%3E72040139%3C/proquest_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=72040139&rft_id=info:pmid/12202241&rfr_iscdi=true |