| Summary: | Urease, which hydrolyzes urea into carbonic acid and ammonia, requires nickel, GTP, bicarbonate, and several accessory proteins for its activation through metallation. This work focuses on the soluble accessory protein complex containing a maltose binding protein/UreD fusion (MBP‐UreD), UreF, and UreG of Klebsiella aerogenes, and its interaction with the cognate urease. The properties of this complex were examined using gel permeation chromatography along with chemical cross‐linking and mass spectrometry of tryptic peptides, revealing it to be an overall dimeric species. Variants of UreD within the complex were analyzed for their ability to bind urease and facilitate its activation. Select UreD variants lacking the maltose binding protein solubility tag were analyzed for their ability to restore urease activation in a ∆ureD urease gene cluster. Also presented are biochemical studies that give further insight into the processes behind urease activation. Grant Funding Source : DK045686
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