Assignments of backbone 1H, 13C and 15N resonances in H-Ras (1–166) complexed with GppNHp at physiological pH
The small GTPase Ras is an important signaling molecule acting as a molecular switch in eukaryotic cells. Recent findings of global conformational exchange and a putative allosteric binding site in the G domain of Ras opened an avenue to understanding novel aspects of Ras function. To facilitate detailed NMR studies of Ras in physiological solution conditions, we performed backbone resonance assignments of Ras bound to slowly hydrolysable GTP mimic, guanosine 5′-[ß, γ-imido]triphosphate at pH 7.2. Out of 163 non-proline residues of the G domain, signals from backbone amide proton, nitrogen and carbon spins of 127 residues were confidently assigned with the remaining unassigned residues mostly located at the exchange-broadened effectors interface.
KeywordsNMR Ras Assignments
Guanosine 5′-[ß, γ-imido]triphosphate
The authors acknowledge Dr. Elizaveta Kovrigina for help with NMR sample preparation and Dr. Francis Peterson for assistance with setting up the triple-resonance NMR experiments. The funding for this work came from the Medical College of Wisconsin as Program Development funds to Evgenii L. Kovrigin.
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