title: The conformational response to Zn(II) and Ni(II) binding of Sporosarcina pasteurii UreG, an intrinsically disordered GTPase.
abstract: Urease is an essential Ni(II) enzyme involved in the nitrogen metabolism of bacteria, plants and fungi. Ni(II) delivery into the enzyme active site requires the presence of four accessory proteins, named UreD, UreF, UreG and UreE, acting through a complex protein network regulated by metal binding and GTP hydrolysis. The GTPase activity is catalyzed by UreG, which couples this function to a non-enzymatic role as a molecular chaperone. This moonlighting activity is reflected in a flexible fold that makes UreG the first discovered intrinsically disordered enzyme. UreG binds Ni(II) and Zn(II),which in turn modulate the interactions with other urease chaperones. The aim of this study is to understand the structural implications of metal binding to Sporosarcina pasteurii UreG (SpUreG). A combination of light scattering, calorimetry, mass spectrometry, and NMR spectroscopy revealed that SpUreG exists in monomer-dimer equilibrium (K(d)= 45 µM), sampling three distinct folding populations with different degrees of compactness. Binding of Zn(II) ions, occurring in two distinct sites (K(d1) = 3 nM, K(d2) = 0.53 µM), shifts the protein conformational landscape toward the more compact population, while maintaining the overall protein structural plasticity. Differently, binding of Ni(II) ions occurs in three binding sites (K(d1(= 14 µM; K(d2) = 270 µM; K(d3)= 160 µM), with much weaker influence on the protein conformational equilibrium. These distinct conformational responses of SpUreG to Ni(II) and Zn(II) binding suggest that selective metal binding modulates protein plasticity, possibly having an impact on the protein-protein interactions and the enzymatic activity of UreG.
authors.affiliation: Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: D'Urzo
authors.forename: Carlo
authors.initials: C
authors.lastname: Santambrogio
authors.forename: Rita
authors.initials: R
authors.lastname: Grandori
authors.forename: Stefano
authors.initials: S
authors.lastname: Ciurli
authors.forename: Barbara
authors.initials: B
authors.lastname: Zambelli
mesh_terms: D001665:Binding Sites; D008968:Molecular Conformation / Q000187:drug effects*; D009532:Nickel / Q000378:metabolism / Q000494:pharmacology*; D058644:Sporosarcina / Q000201:enzymology*; D014510:Urease / Q000737:chemistry* / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D009532:Nickel; D014510:Urease; D015032:Zinc