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Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR.

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.

SUBMITTER: Peters F

PROVIDER: S-EPMC3193991 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature

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Cys-Ph-TAHA: a lanthanide binding tag for RDC and PCS enhanced protein NMR.

Peters Fabian F Maestre-Martinez Mitcheell M Leonov Andrei A Kovačič Lidija L Becker Stefan S Boelens Rolf R Griesinger Christian C

Journal of biomolecular NMR 20110904 3

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 bio ...[more]

PMID: 21892794

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