Protein Structural Studies by Paramagnetic Solid-State NMR Spectroscopy Aided by a Compact Cyclen-Type Cu(II) Binding Tag
Identifieur interne : 001910 ( Main/Exploration ); précédent : 001909; suivant : 001911Protein Structural Studies by Paramagnetic Solid-State NMR Spectroscopy Aided by a Compact Cyclen-Type Cu(II) Binding Tag
Auteurs : Ishita Sengupta [États-Unis] ; Min Gao [États-Unis] ; Rajith J. Arachchige [États-Unis] ; Philippe S. Nadaud [États-Unis] ; Timothy F. Cunningham [États-Unis] ; Sunil Saxena [États-Unis] ; Charles D. Schwieters [États-Unis] ; Christopher P. Jaroniec [États-Unis]Source :
- Journal of biomolecular NMR [ 0925-2738 ] ; 2014.
Abstract
Paramagnetic relaxation enhancements (PREs) are a rich source of structural information in protein solid-state NMR spectroscopy. Here we demonstrate that PRE measurements in natively diamagnetic proteins are facilitated by a thiol-reactive compact, cyclen-based, high-affinity Cu2+ binding tag, 1-(2-(pyridin-2-yldisulfanyl)ethyl)-1,4,7,10-tetraazacyclododecane (TETAC), that overcomes the key shortcomings associated with the use of larger, more flexible metal-binding tags. Using the TETAC-Cu2+ K28C mutant of B1 immunoglobulin-binding domain of protein G as a model, we find that amino acid residues located within ~10 Å of the Cu2+ center experience considerable transverse PREs leading to severely attenuated resonances in 2D 15N-13C correlation spectra. For more distant residues, electron-nucleus distances are accessible via quantitative measurements of longitudinal PREs, and we demonstrate such measurements for 15N-Cu2+ distances up to ~20 Å.
Url:
DOI: 10.1007/s10858-014-9880-9
PubMed: 25432438
PubMed Central: 4304965
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Pmc, to step Corpus: 001773
- to stream Pmc, to step Curation: 001748
- to stream Pmc, to step Checkpoint: 000874
- to stream Ncbi, to step Merge: 004003
- to stream Ncbi, to step Curation: 004003
- to stream Ncbi, to step Checkpoint: 004003
- to stream Main, to step Merge: 001966
- to stream Main, to step Curation: 001910
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Protein Structural Studies by Paramagnetic Solid-State NMR Spectroscopy Aided by a Compact Cyclen-Type Cu(II) Binding Tag</title><author><name sortKey="Sengupta, Ishita" sort="Sengupta, Ishita" uniqKey="Sengupta I" first="Ishita" last="Sengupta">Ishita Sengupta</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Gao, Min" sort="Gao, Min" uniqKey="Gao M" first="Min" last="Gao">Min Gao</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Arachchige, Rajith J" sort="Arachchige, Rajith J" uniqKey="Arachchige R" first="Rajith J." last="Arachchige">Rajith J. Arachchige</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Nadaud, Philippe S" sort="Nadaud, Philippe S" uniqKey="Nadaud P" first="Philippe S." last="Nadaud">Philippe S. Nadaud</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Cunningham, Timothy F" sort="Cunningham, Timothy F" uniqKey="Cunningham T" first="Timothy F." last="Cunningham">Timothy F. Cunningham</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260</wicri:regionArea><orgName type="university">Université de Pittsburgh</orgName><placeName><settlement type="city">Pittsburgh</settlement><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Saxena, Sunil" sort="Saxena, Sunil" uniqKey="Saxena S" first="Sunil" last="Saxena">Sunil Saxena</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260</wicri:regionArea><orgName type="university">Université de Pittsburgh</orgName><placeName><settlement type="city">Pittsburgh</settlement><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Schwieters, Charles D" sort="Schwieters, Charles D" uniqKey="Schwieters C" first="Charles D." last="Schwieters">Charles D. Schwieters</name><affiliation wicri:level="1"><nlm:aff id="A3">Center for Information Technology, National Institutes of Health, Bethesda, Maryland 20892, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Information Technology, National Institutes of Health, Bethesda, Maryland 20892</wicri:regionArea><wicri:noRegion>Maryland 20892</wicri:noRegion></affiliation></author><author><name sortKey="Jaroniec, Christopher P" sort="Jaroniec, Christopher P" uniqKey="Jaroniec C" first="Christopher P." last="Jaroniec">Christopher P. Jaroniec</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25432438</idno><idno type="pmc">4304965</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4304965</idno><idno type="RBID">PMC:4304965</idno><idno type="doi">10.1007/s10858-014-9880-9</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">001773</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001773</idno><idno type="wicri:Area/Pmc/Curation">001748</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">001748</idno><idno type="wicri:Area/Pmc/Checkpoint">000874</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000874</idno><idno type="wicri:Area/Ncbi/Merge">004003</idno><idno type="wicri:Area/Ncbi/Curation">004003</idno><idno type="wicri:Area/Ncbi/Checkpoint">004003</idno><idno type="wicri:doubleKey">0925-2738:2014:Sengupta I:protein:structural:studies</idno><idno type="wicri:Area/Main/Merge">001966</idno><idno type="wicri:Area/Main/Curation">001910</idno><idno type="wicri:Area/Main/Exploration">001910</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Protein Structural Studies by Paramagnetic Solid-State NMR Spectroscopy Aided by a Compact Cyclen-Type Cu(II) Binding Tag</title><author><name sortKey="Sengupta, Ishita" sort="Sengupta, Ishita" uniqKey="Sengupta I" first="Ishita" last="Sengupta">Ishita Sengupta</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Gao, Min" sort="Gao, Min" uniqKey="Gao M" first="Min" last="Gao">Min Gao</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Arachchige, Rajith J" sort="Arachchige, Rajith J" uniqKey="Arachchige R" first="Rajith J." last="Arachchige">Rajith J. Arachchige</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Nadaud, Philippe S" sort="Nadaud, Philippe S" uniqKey="Nadaud P" first="Philippe S." last="Nadaud">Philippe S. Nadaud</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author><author><name sortKey="Cunningham, Timothy F" sort="Cunningham, Timothy F" uniqKey="Cunningham T" first="Timothy F." last="Cunningham">Timothy F. Cunningham</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260</wicri:regionArea><orgName type="university">Université de Pittsburgh</orgName><placeName><settlement type="city">Pittsburgh</settlement><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Saxena, Sunil" sort="Saxena, Sunil" uniqKey="Saxena S" first="Sunil" last="Saxena">Sunil Saxena</name><affiliation wicri:level="4"><nlm:aff id="A2">Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry, University of Pittsburgh, Pittsburgh, Pennsylvania 15260</wicri:regionArea><orgName type="university">Université de Pittsburgh</orgName><placeName><settlement type="city">Pittsburgh</settlement><region type="state">Pennsylvanie</region></placeName></affiliation></author><author><name sortKey="Schwieters, Charles D" sort="Schwieters, Charles D" uniqKey="Schwieters C" first="Charles D." last="Schwieters">Charles D. Schwieters</name><affiliation wicri:level="1"><nlm:aff id="A3">Center for Information Technology, National Institutes of Health, Bethesda, Maryland 20892, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Center for Information Technology, National Institutes of Health, Bethesda, Maryland 20892</wicri:regionArea><wicri:noRegion>Maryland 20892</wicri:noRegion></affiliation></author><author><name sortKey="Jaroniec, Christopher P" sort="Jaroniec, Christopher P" uniqKey="Jaroniec C" first="Christopher P." last="Jaroniec">Christopher P. Jaroniec</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210, United States</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Chemistry and Biochemistry, The Ohio State University, Columbus, Ohio 43210</wicri:regionArea><wicri:noRegion>Ohio 43210</wicri:noRegion></affiliation></author></analytic><series><title level="j">Journal of biomolecular NMR</title><idno type="ISSN">0925-2738</idno><idno type="eISSN">1573-5001</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="P1">Paramagnetic relaxation enhancements (PREs) are a rich source of structural information in protein solid-state NMR spectroscopy. Here we demonstrate that PRE measurements in natively diamagnetic proteins are facilitated by a thiol-reactive compact, cyclen-based, high-affinity Cu<sup>2+</sup> binding tag, 1-(2-(pyridin-2-yldisulfanyl)ethyl)-1,4,7,10-tetraazacyclododecane (TETAC), that overcomes the key shortcomings associated with the use of larger, more flexible metal-binding tags. Using the TETAC-Cu<sup>2+</sup> K28C mutant of B1 immunoglobulin-binding domain of protein G as a model, we find that amino acid residues located within ~10 Å of the Cu<sup>2+</sup> center experience considerable transverse PREs leading to severely attenuated resonances in 2D <sup>15</sup>N-<sup>13</sup>C correlation spectra. For more distant residues, electron-nucleus distances are accessible via quantitative measurements of longitudinal PREs, and we demonstrate such measurements for <sup>15</sup>N-Cu<sup>2+</sup> distances up to ~20 Å.</p></div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Pennsylvanie</li></region><settlement><li>Pittsburgh</li></settlement><orgName><li>Université de Pittsburgh</li></orgName></list><tree><country name="États-Unis"><noRegion><name sortKey="Sengupta, Ishita" sort="Sengupta, Ishita" uniqKey="Sengupta I" first="Ishita" last="Sengupta">Ishita Sengupta</name></noRegion><name sortKey="Arachchige, Rajith J" sort="Arachchige, Rajith J" uniqKey="Arachchige R" first="Rajith J." last="Arachchige">Rajith J. Arachchige</name><name sortKey="Cunningham, Timothy F" sort="Cunningham, Timothy F" uniqKey="Cunningham T" first="Timothy F." last="Cunningham">Timothy F. Cunningham</name><name sortKey="Gao, Min" sort="Gao, Min" uniqKey="Gao M" first="Min" last="Gao">Min Gao</name><name sortKey="Jaroniec, Christopher P" sort="Jaroniec, Christopher P" uniqKey="Jaroniec C" first="Christopher P." last="Jaroniec">Christopher P. Jaroniec</name><name sortKey="Nadaud, Philippe S" sort="Nadaud, Philippe S" uniqKey="Nadaud P" first="Philippe S." last="Nadaud">Philippe S. Nadaud</name><name sortKey="Saxena, Sunil" sort="Saxena, Sunil" uniqKey="Saxena S" first="Sunil" last="Saxena">Sunil Saxena</name><name sortKey="Schwieters, Charles D" sort="Schwieters, Charles D" uniqKey="Schwieters C" first="Charles D." last="Schwieters">Charles D. Schwieters</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Wicri/Amérique/explor/PittsburghV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001910 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 001910 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Wicri/Amérique
|area= PittsburghV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= PMC:4304965
|texte= Protein Structural Studies by Paramagnetic Solid-State NMR Spectroscopy Aided by a Compact Cyclen-Type Cu(II) Binding Tag
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:25432438" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a PittsburghV1
| This area was generated with Dilib version V0.6.38. |  |