Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR
Identifieur interne : 000A03 ( Main/Exploration ); précédent : 000A02; suivant : 000A04Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR
Auteurs : Fabian Peters [Allemagne] ; Mitcheell Maestre-Martinez [Allemagne] ; Andrei Leonov [Allemagne] ; Lidija Kova I [Pays-Bas, Slovénie] ; Stefan Becker [Allemagne] ; Rolf Boelens [Pays-Bas] ; Christian Griesinger [Allemagne]Source :
- Journal of Biomolecular Nmr [ 0925-2738 ] ; 2011.
Abstract
Here we present Cys-Ph-TAHA, a new nonadentate lanthanide tag for the paramagnetic labelling of proteins. The tag can be easily synthesized and is stereochemically homogenous over a wide range of temperatures, yielding NMR spectra with a single set of peaks. Bound to ubiquitin, it induced large residual dipolar couplings and pseudocontact shifts that could be measured easily and agreed very well with the protein structure. We show that Cys-Ph-TAHA can be used to label large proteins that are biochemically challenging such as the Lac repressor in a 90 kDa ternary complex with DNA and inducer.
The online version of this article (doi:10.1007/s10858-011-9560-y) contains supplementary material, which is available to authorized users.
Url:
DOI: 10.1007/s10858-011-9560-y
PubMed: 21892794
PubMed Central: 3193991
Affiliations:
- Allemagne, Pays-Bas, Slovénie
- Basse-Saxe, Utrecht (province)
- Göttingen, Utrecht
- Université d'Utrecht
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Biomolecular Nmr</title><idno type="ISSN">0925-2738</idno><idno type="eISSN">1573-5001</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Here we present Cys-Ph-TAHA, a new nonadentate lanthanide tag for the paramagnetic labelling of proteins. The tag can be easily synthesized and is stereochemically homogenous over a wide range of temperatures, yielding NMR spectra with a single set of peaks. Bound to ubiquitin, it induced large residual dipolar couplings and pseudocontact shifts that could be measured easily and agreed very well with the protein structure. We show that Cys-Ph-TAHA can be used to label large proteins that are biochemically challenging such as the Lac repressor in a 90 kDa ternary complex with DNA and inducer.</p><sec><title>Electronic supplementary material</title><p>The online version of this article (doi:10.1007/s10858-011-9560-y) contains supplementary material, which is available to authorized users.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Allegrozzi, M" uniqKey="Allegrozzi M">M Allegrozzi</name></author><author><name sortKey="Bertini, I" uniqKey="Bertini I">I Bertini</name></author><author><name sortKey="Janik, Mbl" uniqKey="Janik M">MBL Janik</name></author><author><name sortKey="Lee, G" uniqKey="Lee G">G Lee</name></author><author><name sortKey="Liu, Ym" uniqKey="Liu Y">YM Liu</name></author><author><name sortKey="Luchinat, C" uniqKey="Luchinat C">C Luchinat</name></author></analytic></biblStruct><biblStruct><analytic><author><name 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Hannemann</name></author><author><name sortKey="Bernhardt, R" uniqKey="Bernhardt R">R Bernhardt</name></author><author><name sortKey="Ubbink, M" uniqKey="Ubbink M">M Ubbink</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="You, J" uniqKey="You J">J You</name></author><author><name sortKey="Cohen, Re" uniqKey="Cohen R">RE Cohen</name></author><author><name sortKey="Pickart, Cm" uniqKey="Pickart C">CM Pickart</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Allemagne</li><li>Pays-Bas</li><li>Slovénie</li></country><region><li>Basse-Saxe</li><li>Utrecht (province)</li></region><settlement><li>Göttingen</li><li>Utrecht</li></settlement><orgName><li>Université d'Utrecht</li></orgName></list><tree><country name="Allemagne"><region name="Basse-Saxe"><name sortKey="Peters, Fabian" sort="Peters, Fabian" uniqKey="Peters F" first="Fabian" last="Peters">Fabian Peters</name></region><name sortKey="Becker, Stefan" sort="Becker, Stefan" uniqKey="Becker S" first="Stefan" last="Becker">Stefan Becker</name><name sortKey="Griesinger, Christian" sort="Griesinger, Christian" uniqKey="Griesinger C" first="Christian" last="Griesinger">Christian Griesinger</name><name sortKey="Leonov, Andrei" sort="Leonov, Andrei" uniqKey="Leonov A" first="Andrei" last="Leonov">Andrei Leonov</name><name sortKey="Maestre Martinez, Mitcheell" sort="Maestre Martinez, Mitcheell" uniqKey="Maestre Martinez M" first="Mitcheell" last="Maestre-Martinez">Mitcheell Maestre-Martinez</name></country><country name="Pays-Bas"><region name="Utrecht (province)"><name sortKey="Kova I, Lidija" sort="Kova I, Lidija" uniqKey="Kova I L" first="Lidija" last="Kova I">Lidija Kova I</name></region><name sortKey="Boelens, Rolf" sort="Boelens, Rolf" uniqKey="Boelens R" first="Rolf" last="Boelens">Rolf Boelens</name></country><country name="Slovénie"><noRegion><name sortKey="Kova I, Lidija" sort="Kova I, Lidija" uniqKey="Kova I L" 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