title: 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.
authors: Department of NMR-Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
authors: Fabian
authors: F
authors: Peters
authors: Mitcheell
authors: M
authors: Maestre-Martinez
authors: Andrei
authors: A
authors: Leonov
authors: Lidija
authors: L
authors: Kovačič
authors: Stefan
authors: S
authors: Becker
authors: Rolf
authors: R
authors: Boelens
authors: Christian
authors: C
authors: Griesinger
mesh_terms: D000085:Acetates / Q000737:chemistry*; D001665:Binding Sites; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry*; D028581:Lanthanoid Series Elements / Q000737:chemistry*; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D025801:Ubiquitin / Q000737:chemistry
chemical_list: D000085:Acetates; D028581:Lanthanoid Series Elements; D011506:Proteins; D025801:Ubiquitin; C571507:cysteinyl-phenyl-triaminohexaacetate; D003545:Cysteine