Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA
The blue light using flavin (BLUF) domain proteins, such as the transcriptional antirepressor AppA, are a novel class of photosensors that bind flavin noncovalently in order to sense and respond to high-intensity blue (450 nm) light. Importantly, the noncovalently bound flavin chromophore is unable...
Gespeichert in:
| Veröffentlicht in: | The journal of physical chemistry. B 2012-09, Vol.116 (35), p.10722-10729 |
|---|---|
| Hauptverfasser: | , , , , , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 10729 |
|---|---|
| container_issue | 35 |
| container_start_page | 10722 |
| container_title | The journal of physical chemistry. B |
| container_volume | 116 |
| creator | Haigney, Allison Lukacs, Andras Brust, Richard Zhao, Rui-Kun Towrie, Michael Greetham, Gregory M Clark, Ian Illarionov, Boris Bacher, Adelbert Kim, Ryu-Ryun Fischer, Markus Meech, Stephen R Tonge, Peter J |
| description | The blue light using flavin (BLUF) domain proteins, such as the transcriptional antirepressor AppA, are a novel class of photosensors that bind flavin noncovalently in order to sense and respond to high-intensity blue (450 nm) light. Importantly, the noncovalently bound flavin chromophore is unable to undergo large-scale structural change upon light absorption, and thus there is significant interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppABLUF) reconstituted with isotopes of FAD, specifically [U-13C17]-FAD, [xylene-13C8]-FAD, [U-15N4]-FAD, and [4-18O1]-FAD both in solution and bound to AppABLUF. This allows for unambiguous assignment of ground- and excited-state modes arising directly from the flavin. Studies of model compounds and DFT calculations of the ground-state vibrational spectra reveal the sensitivity of these modes to their environment, indicating they can be used as probes of structural dynamics. |
| doi_str_mv | 10.1021/jp305220m |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_1753475048</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1753475048</sourcerecordid><originalsourceid>FETCH-LOGICAL-a414t-9f4b76fa1c147576d666c05e0d6ea13e8e0c0d8c61bceade97271df58991ab023</originalsourceid><addsrcrecordid>eNqF0EFLwzAUB_AgipvTg19AchH0UE3SNm2PYzgdDBR0Xstr-uo62qYm6cBvb2SbJ8FDeOHx4__gT8glZ3ecCX6_6UMWC8HaIzLmsWCBf8nx_i85kyNyZu2GMRGLVJ6SkRBp4tdyTNR7XRhwte6goVNr64-uxc5RXVG3RrpqnIEKrKOLrjJgsKSvPSpnhvZAXtbaaVCu3oKDokE6b2Cre6Md1h2d9v30nJxU0Fi82M8JWc0f3mZPwfL5cTGbLgOIeOSCrIqKRFbAFY-SOJGllFKxGFkpEXiIKTLFylRJXiiEErNEJLys4jTLOBRMhBNys8v1xz8HtC5va6uwaaBDPdicJ3Hok1mU_k9ZmMZZJCPm6e2OKqOtNVjlvalbMF8e5T_157_1e3u1jx2KFstfeejbg-sdAGXzjR6Mr93-EfQN2QeMSw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1038594640</pqid></control><display><type>article</type><title>Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA</title><source>MEDLINE</source><source>American Chemical Society Journals</source><creator>Haigney, Allison ; Lukacs, Andras ; Brust, Richard ; Zhao, Rui-Kun ; Towrie, Michael ; Greetham, Gregory M ; Clark, Ian ; Illarionov, Boris ; Bacher, Adelbert ; Kim, Ryu-Ryun ; Fischer, Markus ; Meech, Stephen R ; Tonge, Peter J</creator><creatorcontrib>Haigney, Allison ; Lukacs, Andras ; Brust, Richard ; Zhao, Rui-Kun ; Towrie, Michael ; Greetham, Gregory M ; Clark, Ian ; Illarionov, Boris ; Bacher, Adelbert ; Kim, Ryu-Ryun ; Fischer, Markus ; Meech, Stephen R ; Tonge, Peter J</creatorcontrib><description>The blue light using flavin (BLUF) domain proteins, such as the transcriptional antirepressor AppA, are a novel class of photosensors that bind flavin noncovalently in order to sense and respond to high-intensity blue (450 nm) light. Importantly, the noncovalently bound flavin chromophore is unable to undergo large-scale structural change upon light absorption, and thus there is significant interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppABLUF) reconstituted with isotopes of FAD, specifically [U-13C17]-FAD, [xylene-13C8]-FAD, [U-15N4]-FAD, and [4-18O1]-FAD both in solution and bound to AppABLUF. This allows for unambiguous assignment of ground- and excited-state modes arising directly from the flavin. Studies of model compounds and DFT calculations of the ground-state vibrational spectra reveal the sensitivity of these modes to their environment, indicating they can be used as probes of structural dynamics.</description><identifier>ISSN: 1520-6106</identifier><identifier>EISSN: 1520-5207</identifier><identifier>DOI: 10.1021/jp305220m</identifier><identifier>PMID: 22871066</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Bacterial Proteins - chemistry ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; Chromophores ; Dynamic structural analysis ; Flavoproteins - chemistry ; Flavoproteins - genetics ; Flavoproteins - metabolism ; Hydrogen Bonding ; Light absorption ; Mathematical models ; Networks ; Photoexcitation ; Protein Structure, Tertiary ; Proteins ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Rhodobacter sphaeroides - metabolism ; Spectrophotometry, Infrared ; Vibration</subject><ispartof>The journal of physical chemistry. B, 2012-09, Vol.116 (35), p.10722-10729</ispartof><rights>Copyright © 2012 American Chemical Society</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a414t-9f4b76fa1c147576d666c05e0d6ea13e8e0c0d8c61bceade97271df58991ab023</citedby><cites>FETCH-LOGICAL-a414t-9f4b76fa1c147576d666c05e0d6ea13e8e0c0d8c61bceade97271df58991ab023</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/jp305220m$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/jp305220m$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,776,780,2752,27053,27901,27902,56713,56763</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22871066$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Haigney, Allison</creatorcontrib><creatorcontrib>Lukacs, Andras</creatorcontrib><creatorcontrib>Brust, Richard</creatorcontrib><creatorcontrib>Zhao, Rui-Kun</creatorcontrib><creatorcontrib>Towrie, Michael</creatorcontrib><creatorcontrib>Greetham, Gregory M</creatorcontrib><creatorcontrib>Clark, Ian</creatorcontrib><creatorcontrib>Illarionov, Boris</creatorcontrib><creatorcontrib>Bacher, Adelbert</creatorcontrib><creatorcontrib>Kim, Ryu-Ryun</creatorcontrib><creatorcontrib>Fischer, Markus</creatorcontrib><creatorcontrib>Meech, Stephen R</creatorcontrib><creatorcontrib>Tonge, Peter J</creatorcontrib><title>Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA</title><title>The journal of physical chemistry. B</title><addtitle>J. Phys. Chem. B</addtitle><description>The blue light using flavin (BLUF) domain proteins, such as the transcriptional antirepressor AppA, are a novel class of photosensors that bind flavin noncovalently in order to sense and respond to high-intensity blue (450 nm) light. Importantly, the noncovalently bound flavin chromophore is unable to undergo large-scale structural change upon light absorption, and thus there is significant interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppABLUF) reconstituted with isotopes of FAD, specifically [U-13C17]-FAD, [xylene-13C8]-FAD, [U-15N4]-FAD, and [4-18O1]-FAD both in solution and bound to AppABLUF. This allows for unambiguous assignment of ground- and excited-state modes arising directly from the flavin. Studies of model compounds and DFT calculations of the ground-state vibrational spectra reveal the sensitivity of these modes to their environment, indicating they can be used as probes of structural dynamics.</description><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>Chromophores</subject><subject>Dynamic structural analysis</subject><subject>Flavoproteins - chemistry</subject><subject>Flavoproteins - genetics</subject><subject>Flavoproteins - metabolism</subject><subject>Hydrogen Bonding</subject><subject>Light absorption</subject><subject>Mathematical models</subject><subject>Networks</subject><subject>Photoexcitation</subject><subject>Protein Structure, Tertiary</subject><subject>Proteins</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Rhodobacter sphaeroides - metabolism</subject><subject>Spectrophotometry, Infrared</subject><subject>Vibration</subject><issn>1520-6106</issn><issn>1520-5207</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0EFLwzAUB_AgipvTg19AchH0UE3SNm2PYzgdDBR0Xstr-uo62qYm6cBvb2SbJ8FDeOHx4__gT8glZ3ecCX6_6UMWC8HaIzLmsWCBf8nx_i85kyNyZu2GMRGLVJ6SkRBp4tdyTNR7XRhwte6goVNr64-uxc5RXVG3RrpqnIEKrKOLrjJgsKSvPSpnhvZAXtbaaVCu3oKDokE6b2Cre6Md1h2d9v30nJxU0Fi82M8JWc0f3mZPwfL5cTGbLgOIeOSCrIqKRFbAFY-SOJGllFKxGFkpEXiIKTLFylRJXiiEErNEJLys4jTLOBRMhBNys8v1xz8HtC5va6uwaaBDPdicJ3Hok1mU_k9ZmMZZJCPm6e2OKqOtNVjlvalbMF8e5T_157_1e3u1jx2KFstfeejbg-sdAGXzjR6Mr93-EfQN2QeMSw</recordid><startdate>20120906</startdate><enddate>20120906</enddate><creator>Haigney, Allison</creator><creator>Lukacs, Andras</creator><creator>Brust, Richard</creator><creator>Zhao, Rui-Kun</creator><creator>Towrie, Michael</creator><creator>Greetham, Gregory M</creator><creator>Clark, Ian</creator><creator>Illarionov, Boris</creator><creator>Bacher, Adelbert</creator><creator>Kim, Ryu-Ryun</creator><creator>Fischer, Markus</creator><creator>Meech, Stephen R</creator><creator>Tonge, Peter J</creator><general>American Chemical Society</general><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><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope></search><sort><creationdate>20120906</creationdate><title>Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA</title><author>Haigney, Allison ; Lukacs, Andras ; Brust, Richard ; Zhao, Rui-Kun ; Towrie, Michael ; Greetham, Gregory M ; Clark, Ian ; Illarionov, Boris ; Bacher, Adelbert ; Kim, Ryu-Ryun ; Fischer, Markus ; Meech, Stephen R ; Tonge, Peter J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a414t-9f4b76fa1c147576d666c05e0d6ea13e8e0c0d8c61bceade97271df58991ab023</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>Chromophores</topic><topic>Dynamic structural analysis</topic><topic>Flavoproteins - chemistry</topic><topic>Flavoproteins - genetics</topic><topic>Flavoproteins - metabolism</topic><topic>Hydrogen Bonding</topic><topic>Light absorption</topic><topic>Mathematical models</topic><topic>Networks</topic><topic>Photoexcitation</topic><topic>Protein Structure, Tertiary</topic><topic>Proteins</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Recombinant Proteins - metabolism</topic><topic>Rhodobacter sphaeroides - metabolism</topic><topic>Spectrophotometry, Infrared</topic><topic>Vibration</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Haigney, Allison</creatorcontrib><creatorcontrib>Lukacs, Andras</creatorcontrib><creatorcontrib>Brust, Richard</creatorcontrib><creatorcontrib>Zhao, Rui-Kun</creatorcontrib><creatorcontrib>Towrie, Michael</creatorcontrib><creatorcontrib>Greetham, Gregory M</creatorcontrib><creatorcontrib>Clark, Ian</creatorcontrib><creatorcontrib>Illarionov, Boris</creatorcontrib><creatorcontrib>Bacher, Adelbert</creatorcontrib><creatorcontrib>Kim, Ryu-Ryun</creatorcontrib><creatorcontrib>Fischer, Markus</creatorcontrib><creatorcontrib>Meech, Stephen R</creatorcontrib><creatorcontrib>Tonge, Peter J</creatorcontrib><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><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><jtitle>The journal of physical chemistry. B</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Haigney, Allison</au><au>Lukacs, Andras</au><au>Brust, Richard</au><au>Zhao, Rui-Kun</au><au>Towrie, Michael</au><au>Greetham, Gregory M</au><au>Clark, Ian</au><au>Illarionov, Boris</au><au>Bacher, Adelbert</au><au>Kim, Ryu-Ryun</au><au>Fischer, Markus</au><au>Meech, Stephen R</au><au>Tonge, Peter J</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA</atitle><jtitle>The journal of physical chemistry. B</jtitle><addtitle>J. Phys. Chem. B</addtitle><date>2012-09-06</date><risdate>2012</risdate><volume>116</volume><issue>35</issue><spage>10722</spage><epage>10729</epage><pages>10722-10729</pages><issn>1520-6106</issn><eissn>1520-5207</eissn><abstract>The blue light using flavin (BLUF) domain proteins, such as the transcriptional antirepressor AppA, are a novel class of photosensors that bind flavin noncovalently in order to sense and respond to high-intensity blue (450 nm) light. Importantly, the noncovalently bound flavin chromophore is unable to undergo large-scale structural change upon light absorption, and thus there is significant interest in understanding how the BLUF protein matrix senses and responds to flavin photoexcitation. Light absorption is proposed to result in alterations in the hydrogen-bonding network that surrounds the flavin chromophore on an ultrafast time scale, and the structural changes caused by photoexcitation are being probed by vibrational spectroscopy. Here we report ultrafast time-resolved infrared spectra of the AppA BLUF domain (AppABLUF) reconstituted with isotopes of FAD, specifically [U-13C17]-FAD, [xylene-13C8]-FAD, [U-15N4]-FAD, and [4-18O1]-FAD both in solution and bound to AppABLUF. This allows for unambiguous assignment of ground- and excited-state modes arising directly from the flavin. Studies of model compounds and DFT calculations of the ground-state vibrational spectra reveal the sensitivity of these modes to their environment, indicating they can be used as probes of structural dynamics.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>22871066</pmid><doi>10.1021/jp305220m</doi><tpages>8</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1520-6106 |
| ispartof | The journal of physical chemistry. B, 2012-09, Vol.116 (35), p.10722-10729 |
| issn | 1520-6106 1520-5207 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1753475048 |
| source | MEDLINE; American Chemical Society Journals |
| subjects | Bacterial Proteins - chemistry Bacterial Proteins - genetics Bacterial Proteins - metabolism Chromophores Dynamic structural analysis Flavoproteins - chemistry Flavoproteins - genetics Flavoproteins - metabolism Hydrogen Bonding Light absorption Mathematical models Networks Photoexcitation Protein Structure, Tertiary Proteins Recombinant Proteins - chemistry Recombinant Proteins - genetics Recombinant Proteins - metabolism Rhodobacter sphaeroides - metabolism Spectrophotometry, Infrared Vibration |
| title | Vibrational Assignment of the Ultrafast Infrared Spectrum of the Photoactivatable Flavoprotein AppA |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-03T06%3A51%3A02IST&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=Vibrational%20Assignment%20of%20the%20Ultrafast%20Infrared%20Spectrum%20of%20the%20Photoactivatable%20Flavoprotein%20AppA&rft.jtitle=The%20journal%20of%20physical%20chemistry.%20B&rft.au=Haigney,%20Allison&rft.date=2012-09-06&rft.volume=116&rft.issue=35&rft.spage=10722&rft.epage=10729&rft.pages=10722-10729&rft.issn=1520-6106&rft.eissn=1520-5207&rft_id=info:doi/10.1021/jp305220m&rft_dat=%3Cproquest_cross%3E1753475048%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=1038594640&rft_id=info:pmid/22871066&rfr_iscdi=true |