Paramagnetic Solid-state NMR to Localize the Metal-Ion Cofactor in an Oligomeric DnaB Helicase
 

Paramagnetic Solid-state NMR to Localize the Metal-Ion Cofactor in an Oligomeric DnaB Helicase

Paramagnetic metal ions can be inserted into ATP-fueled motor proteins by exchanging the diamagnetic Mg 2+ cofactor with Mn 2+ or Co 2+ . Then, paramagnetic relaxation enhancement (PRE) or pseudo-contact shifts (PCSs) can be measured to report on the localization of the metal ion within the protein. We determine the metal position in the oligomeric bacterial DnaB helicase from Helicobacter pylori complexed with the transition-state ATP-analogue ADP:AlF 4 - and single-stranded DNA using...

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