Loading…
Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain
The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a stru...
Saved in:
Published in: | Faraday discussions 2015-01, Vol.177, p.293-311 |
---|---|
Main Authors: | , , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
cited_by | cdi_FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3 |
---|---|
cites | cdi_FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3 |
container_end_page | 311 |
container_issue | |
container_start_page | 293 |
container_title | Faraday discussions |
container_volume | 177 |
creator | Laptenok, Sergey P Lukacs, Andras Brust, Richard Haigney, Allison Gil, Agnieszka Towrie, Michael Greetham, Gregory M Tonge, Peter J Meech, Stephen R |
description | The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a structure change to form the signalling state of the protein, is unresolved. In this work we apply ultrafast time resolved infra-red (TRIR) spectroscopy to investigate the primary photophysics of the BLUF domain of the protein AppA (AppA
BLUF
) a light activated antirepressor. Here a number of mutations at Y21 and W104 in AppA
BLUF
are investigated. The Y21 mutants are known to be photoinactive, while W104 mutants show the characteristic spectral red-shift associated with BLUF domain activity. Using TRIR we observed separately the decay of the excited state and the recovery of the ground state. In both cases the kinetics are found to be non-single exponential for all the proteins studied, suggesting a range of ground state structures. In the Y21 mutants an intermediate state was also observed, assigned to formation of the radical of the isoalloxazine (flavin) ring. The electron donor is the W104 residue. In contrast, no radical intermediates were detected in the studies of the photoactive dark adapted proteins, dAppA
BLUF
and the dW104 mutants, suggesting a structure change in the Y21 mutants which favours W104 to isoalloxazine electron transfer. In contrast, in the light adapted form of the proteins (lAppA
BLUF
, lW104) a radical intermediate was detected and the kinetics were greatly accelerated. In this case the electron donor was Y21 and major structural changes are associated with the enhanced quenching. In AppA
BLUF
and the seven mutants studied radical intermediates are readily observed by TRIR spectroscopy, but there is no correlation with photoactivity. This suggests that if a charge separated state has a role in the BLUF photocycle it is only as a very short lived intermediate. |
doi_str_mv | 10.1039/c4fd00189c |
format | article |
fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_crossref_primary_10_1039_C4FD00189C</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1673375435</sourcerecordid><originalsourceid>FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3</originalsourceid><addsrcrecordid>eNqF0UtP3DAUBWALgToD7YZ9kbtDlUKv42eWw5QBpJHYMOvI8YNJlTjBdhb99w0MpTu6upbup6OrY4TOCVwRoNUPw7wFIKoyR2hJqGAFZ5U6fnnzqhCCwQKdpvQLAMS8_YQWJReUMgVLtLvpnMlxCDhHHZJ3ET9PLph9G55wG3DXPu0z1laP2Vmsg8X9lHXI2A-xT3jwOO8dXo3jCl9vdxtsh1634TM68bpL7svbPEO7zc3j-q7YPtzer1fbwnCQuaga05RCEqU8cOEUEYyDAlWVghmAhpnSapBSEMOsL6tGci6pKsFKR4h19AxdHnLHOMxnp1z3bTKu63Rww5RqogAYnfPZ_6mQlErOKJ_p9wM1cUgpOl-Pse11_F0TqF8ar9ds8_O18fWML95yp6Z39p3-rXgG3w4gJvO-_fdl9Wj9bL5-ZOgfWVCOFw</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1673375435</pqid></control><display><type>article</type><title>Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain</title><source>Royal Society of Chemistry</source><creator>Laptenok, Sergey P ; Lukacs, Andras ; Brust, Richard ; Haigney, Allison ; Gil, Agnieszka ; Towrie, Michael ; Greetham, Gregory M ; Tonge, Peter J ; Meech, Stephen R</creator><creatorcontrib>Laptenok, Sergey P ; Lukacs, Andras ; Brust, Richard ; Haigney, Allison ; Gil, Agnieszka ; Towrie, Michael ; Greetham, Gregory M ; Tonge, Peter J ; Meech, Stephen R</creatorcontrib><description>The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a structure change to form the signalling state of the protein, is unresolved. In this work we apply ultrafast time resolved infra-red (TRIR) spectroscopy to investigate the primary photophysics of the BLUF domain of the protein AppA (AppA
BLUF
) a light activated antirepressor. Here a number of mutations at Y21 and W104 in AppA
BLUF
are investigated. The Y21 mutants are known to be photoinactive, while W104 mutants show the characteristic spectral red-shift associated with BLUF domain activity. Using TRIR we observed separately the decay of the excited state and the recovery of the ground state. In both cases the kinetics are found to be non-single exponential for all the proteins studied, suggesting a range of ground state structures. In the Y21 mutants an intermediate state was also observed, assigned to formation of the radical of the isoalloxazine (flavin) ring. The electron donor is the W104 residue. In contrast, no radical intermediates were detected in the studies of the photoactive dark adapted proteins, dAppA
BLUF
and the dW104 mutants, suggesting a structure change in the Y21 mutants which favours W104 to isoalloxazine electron transfer. In contrast, in the light adapted form of the proteins (lAppA
BLUF
, lW104) a radical intermediate was detected and the kinetics were greatly accelerated. In this case the electron donor was Y21 and major structural changes are associated with the enhanced quenching. In AppA
BLUF
and the seven mutants studied radical intermediates are readily observed by TRIR spectroscopy, but there is no correlation with photoactivity. This suggests that if a charge separated state has a role in the BLUF photocycle it is only as a very short lived intermediate.</description><identifier>ISSN: 1359-6640</identifier><identifier>EISSN: 1364-5498</identifier><identifier>DOI: 10.1039/c4fd00189c</identifier><identifier>PMID: 25633480</identifier><language>eng</language><publisher>England</publisher><subject>Bacterial Proteins - chemistry ; Bacterial Proteins - genetics ; Charge ; Electron transfer ; Electron Transport ; Electrons ; Escherichia coli - genetics ; Escherichia coli - metabolism ; Flavins - chemistry ; Flavoproteins - chemistry ; Flavoproteins - genetics ; Free Radicals - chemistry ; Gene Expression ; Ground state ; Kinetics ; Light ; Models, Molecular ; Mutation ; Photoreceptors ; Photoreceptors, Microbial - chemistry ; Photoreceptors, Microbial - genetics ; Protein Structure, Tertiary ; Proteins ; Quenching ; Radicals ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Spectroscopy ; Spectroscopy, Fourier Transform Infrared - methods ; Static Electricity ; Structure-Activity Relationship ; Time Factors</subject><ispartof>Faraday discussions, 2015-01, Vol.177, p.293-311</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3</citedby><cites>FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3</cites></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/25633480$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Laptenok, Sergey P</creatorcontrib><creatorcontrib>Lukacs, Andras</creatorcontrib><creatorcontrib>Brust, Richard</creatorcontrib><creatorcontrib>Haigney, Allison</creatorcontrib><creatorcontrib>Gil, Agnieszka</creatorcontrib><creatorcontrib>Towrie, Michael</creatorcontrib><creatorcontrib>Greetham, Gregory M</creatorcontrib><creatorcontrib>Tonge, Peter J</creatorcontrib><creatorcontrib>Meech, Stephen R</creatorcontrib><title>Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain</title><title>Faraday discussions</title><addtitle>Faraday Discuss</addtitle><description>The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a structure change to form the signalling state of the protein, is unresolved. In this work we apply ultrafast time resolved infra-red (TRIR) spectroscopy to investigate the primary photophysics of the BLUF domain of the protein AppA (AppA
BLUF
) a light activated antirepressor. Here a number of mutations at Y21 and W104 in AppA
BLUF
are investigated. The Y21 mutants are known to be photoinactive, while W104 mutants show the characteristic spectral red-shift associated with BLUF domain activity. Using TRIR we observed separately the decay of the excited state and the recovery of the ground state. In both cases the kinetics are found to be non-single exponential for all the proteins studied, suggesting a range of ground state structures. In the Y21 mutants an intermediate state was also observed, assigned to formation of the radical of the isoalloxazine (flavin) ring. The electron donor is the W104 residue. In contrast, no radical intermediates were detected in the studies of the photoactive dark adapted proteins, dAppA
BLUF
and the dW104 mutants, suggesting a structure change in the Y21 mutants which favours W104 to isoalloxazine electron transfer. In contrast, in the light adapted form of the proteins (lAppA
BLUF
, lW104) a radical intermediate was detected and the kinetics were greatly accelerated. In this case the electron donor was Y21 and major structural changes are associated with the enhanced quenching. In AppA
BLUF
and the seven mutants studied radical intermediates are readily observed by TRIR spectroscopy, but there is no correlation with photoactivity. This suggests that if a charge separated state has a role in the BLUF photocycle it is only as a very short lived intermediate.</description><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Proteins - genetics</subject><subject>Charge</subject><subject>Electron transfer</subject><subject>Electron Transport</subject><subject>Electrons</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - metabolism</subject><subject>Flavins - chemistry</subject><subject>Flavoproteins - chemistry</subject><subject>Flavoproteins - genetics</subject><subject>Free Radicals - chemistry</subject><subject>Gene Expression</subject><subject>Ground state</subject><subject>Kinetics</subject><subject>Light</subject><subject>Models, Molecular</subject><subject>Mutation</subject><subject>Photoreceptors</subject><subject>Photoreceptors, Microbial - chemistry</subject><subject>Photoreceptors, Microbial - genetics</subject><subject>Protein Structure, Tertiary</subject><subject>Proteins</subject><subject>Quenching</subject><subject>Radicals</subject><subject>Recombinant Proteins - chemistry</subject><subject>Recombinant Proteins - genetics</subject><subject>Spectroscopy</subject><subject>Spectroscopy, Fourier Transform Infrared - methods</subject><subject>Static Electricity</subject><subject>Structure-Activity Relationship</subject><subject>Time Factors</subject><issn>1359-6640</issn><issn>1364-5498</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNqF0UtP3DAUBWALgToD7YZ9kbtDlUKv42eWw5QBpJHYMOvI8YNJlTjBdhb99w0MpTu6upbup6OrY4TOCVwRoNUPw7wFIKoyR2hJqGAFZ5U6fnnzqhCCwQKdpvQLAMS8_YQWJReUMgVLtLvpnMlxCDhHHZJ3ET9PLph9G55wG3DXPu0z1laP2Vmsg8X9lHXI2A-xT3jwOO8dXo3jCl9vdxtsh1634TM68bpL7svbPEO7zc3j-q7YPtzer1fbwnCQuaga05RCEqU8cOEUEYyDAlWVghmAhpnSapBSEMOsL6tGci6pKsFKR4h19AxdHnLHOMxnp1z3bTKu63Rww5RqogAYnfPZ_6mQlErOKJ_p9wM1cUgpOl-Pse11_F0TqF8ar9ds8_O18fWML95yp6Z39p3-rXgG3w4gJvO-_fdl9Wj9bL5-ZOgfWVCOFw</recordid><startdate>20150101</startdate><enddate>20150101</enddate><creator>Laptenok, Sergey P</creator><creator>Lukacs, Andras</creator><creator>Brust, Richard</creator><creator>Haigney, Allison</creator><creator>Gil, Agnieszka</creator><creator>Towrie, Michael</creator><creator>Greetham, Gregory M</creator><creator>Tonge, Peter J</creator><creator>Meech, Stephen R</creator><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>8BQ</scope><scope>8FD</scope><scope>JG9</scope></search><sort><creationdate>20150101</creationdate><title>Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain</title><author>Laptenok, Sergey P ; Lukacs, Andras ; Brust, Richard ; Haigney, Allison ; Gil, Agnieszka ; Towrie, Michael ; Greetham, Gregory M ; Tonge, Peter J ; Meech, Stephen R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Proteins - genetics</topic><topic>Charge</topic><topic>Electron transfer</topic><topic>Electron Transport</topic><topic>Electrons</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - metabolism</topic><topic>Flavins - chemistry</topic><topic>Flavoproteins - chemistry</topic><topic>Flavoproteins - genetics</topic><topic>Free Radicals - chemistry</topic><topic>Gene Expression</topic><topic>Ground state</topic><topic>Kinetics</topic><topic>Light</topic><topic>Models, Molecular</topic><topic>Mutation</topic><topic>Photoreceptors</topic><topic>Photoreceptors, Microbial - chemistry</topic><topic>Photoreceptors, Microbial - genetics</topic><topic>Protein Structure, Tertiary</topic><topic>Proteins</topic><topic>Quenching</topic><topic>Radicals</topic><topic>Recombinant Proteins - chemistry</topic><topic>Recombinant Proteins - genetics</topic><topic>Spectroscopy</topic><topic>Spectroscopy, Fourier Transform Infrared - methods</topic><topic>Static Electricity</topic><topic>Structure-Activity Relationship</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Laptenok, Sergey P</creatorcontrib><creatorcontrib>Lukacs, Andras</creatorcontrib><creatorcontrib>Brust, Richard</creatorcontrib><creatorcontrib>Haigney, Allison</creatorcontrib><creatorcontrib>Gil, Agnieszka</creatorcontrib><creatorcontrib>Towrie, Michael</creatorcontrib><creatorcontrib>Greetham, Gregory M</creatorcontrib><creatorcontrib>Tonge, Peter J</creatorcontrib><creatorcontrib>Meech, Stephen R</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>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><jtitle>Faraday discussions</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Laptenok, Sergey P</au><au>Lukacs, Andras</au><au>Brust, Richard</au><au>Haigney, Allison</au><au>Gil, Agnieszka</au><au>Towrie, Michael</au><au>Greetham, Gregory M</au><au>Tonge, Peter J</au><au>Meech, Stephen R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain</atitle><jtitle>Faraday discussions</jtitle><addtitle>Faraday Discuss</addtitle><date>2015-01-01</date><risdate>2015</risdate><volume>177</volume><spage>293</spage><epage>311</epage><pages>293-311</pages><issn>1359-6640</issn><eissn>1364-5498</eissn><abstract>The Blue Light Using Flavin (BLUF) domain proteins are an important family of photoreceptors controlling a range of responses in a wide variety of organisms. The details of the primary photochemical mechanism, by which light absorption in the isoalloxazine ring of the flavin is converted into a structure change to form the signalling state of the protein, is unresolved. In this work we apply ultrafast time resolved infra-red (TRIR) spectroscopy to investigate the primary photophysics of the BLUF domain of the protein AppA (AppA
BLUF
) a light activated antirepressor. Here a number of mutations at Y21 and W104 in AppA
BLUF
are investigated. The Y21 mutants are known to be photoinactive, while W104 mutants show the characteristic spectral red-shift associated with BLUF domain activity. Using TRIR we observed separately the decay of the excited state and the recovery of the ground state. In both cases the kinetics are found to be non-single exponential for all the proteins studied, suggesting a range of ground state structures. In the Y21 mutants an intermediate state was also observed, assigned to formation of the radical of the isoalloxazine (flavin) ring. The electron donor is the W104 residue. In contrast, no radical intermediates were detected in the studies of the photoactive dark adapted proteins, dAppA
BLUF
and the dW104 mutants, suggesting a structure change in the Y21 mutants which favours W104 to isoalloxazine electron transfer. In contrast, in the light adapted form of the proteins (lAppA
BLUF
, lW104) a radical intermediate was detected and the kinetics were greatly accelerated. In this case the electron donor was Y21 and major structural changes are associated with the enhanced quenching. In AppA
BLUF
and the seven mutants studied radical intermediates are readily observed by TRIR spectroscopy, but there is no correlation with photoactivity. This suggests that if a charge separated state has a role in the BLUF photocycle it is only as a very short lived intermediate.</abstract><cop>England</cop><pmid>25633480</pmid><doi>10.1039/c4fd00189c</doi><tpages>19</tpages><oa>free_for_read</oa></addata></record> |
fulltext | fulltext |
identifier | ISSN: 1359-6640 |
ispartof | Faraday discussions, 2015-01, Vol.177, p.293-311 |
issn | 1359-6640 1364-5498 |
language | eng |
recordid | cdi_crossref_primary_10_1039_C4FD00189C |
source | Royal Society of Chemistry |
subjects | Bacterial Proteins - chemistry Bacterial Proteins - genetics Charge Electron transfer Electron Transport Electrons Escherichia coli - genetics Escherichia coli - metabolism Flavins - chemistry Flavoproteins - chemistry Flavoproteins - genetics Free Radicals - chemistry Gene Expression Ground state Kinetics Light Models, Molecular Mutation Photoreceptors Photoreceptors, Microbial - chemistry Photoreceptors, Microbial - genetics Protein Structure, Tertiary Proteins Quenching Radicals Recombinant Proteins - chemistry Recombinant Proteins - genetics Spectroscopy Spectroscopy, Fourier Transform Infrared - methods Static Electricity Structure-Activity Relationship Time Factors |
title | Electron transfer quenching in light adapted and mutant forms of the AppA BLUF domain |
url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-24T12%3A41%3A23IST&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=Electron%20transfer%20quenching%20in%20light%20adapted%20and%20mutant%20forms%20of%20the%20AppA%20BLUF%20domain&rft.jtitle=Faraday%20discussions&rft.au=Laptenok,%20Sergey%20P&rft.date=2015-01-01&rft.volume=177&rft.spage=293&rft.epage=311&rft.pages=293-311&rft.issn=1359-6640&rft.eissn=1364-5498&rft_id=info:doi/10.1039/c4fd00189c&rft_dat=%3Cproquest_cross%3E1673375435%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c507t-9bcb267188f056e8164508089264c00b4c2da07761c4df29b75573820d7e11de3%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1673375435&rft_id=info:pmid/25633480&rfr_iscdi=true |