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| - Sequence-Specific and Stereospecific Assignment of MethylGroups Using Paramagnetic Lanthanides
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| - Pseudocontact shifts (PCSs) induced by a site-specifically bound paramagnetic lanthanide ionare shown to provide fast access to sequence-specific resonance assignments of methyl groups in proteinsof known three-dimensional structure. Stereospecific assignments of Val and Leu methyls are obtained aswell as resonance assignments of all other methyls, including Met εCH3 groups. No prior assignments ofthe diamagnetic protein are required nor are experiments that transfer magnetization between the methylgroups and the protein backbone. Methyl Cz-exchange experiments were designed to provide convenientaccess to PCS measurements in situations where a paramagnetic lanthanide is in exchange with adiamagnetic lanthanide. In the absence of exchange, simultaneous 13C-HSQC assignments and PCSmeasurements are delivered by the newly developed program Possum. The approaches are demonstratedwith the complex between the N-terminal domain of the subunit ε and the subunit θ of the Escherichia coliDNA polymerase III.
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