title: Effects of salt and nickel ion on the conformational stability of Bacillus pasteurii UreE.
abstract: UreE, a urease accessory protein, is proposed to be a metallochaperone assisting the nickel incorporation into the urease active site. We investigated the effects of salt and nickel on the conformational stability of the UreE from Bacillus pasteurii (BpUreE), by circular dichroism (CD) and nuclear magnetic resonance spectroscopy accompanying a thermodynamic inspection. Far-UV CD spectra of BpUreE showed that both salt and nickel stabilized the ordered structure of the protein. The thermal denaturing of BpUreE showed a bimodal feature with an aggregation process before thermal unfolding. This thermally induced aggregation could be suppressed by the addition of salt up to 50 mM, and the further addition of salt increased the thermal resistance of the protein. The nickel addition also elevated the thermal resistance of BpUreE, although it could not prevent the aggregating process. Additionally, the stoichiometry of a specific nickel binding to BpUreE was revealed as one nickel per dimer. Altogether, the present results establish a rather detailed characterization of the thermostability and nickel-binding property of BpUreE.
authors.affiliation: Research Institute of Pharmaceutical Science, College of Pharmacy, Seoul National University, San 56-1, Shinlim-Dong, Kwanak-Gu, South Korea.
authors.forename: Yeon-Hee
authors.initials: YH
authors.lastname: Lee
authors.forename: Hyung-Sik
authors.initials: HS
authors.lastname: Won
authors.forename: Mann-Hyung
authors.initials: MH
authors.lastname: Lee
authors.forename: Bong-Jin
authors.initials: BJ
authors.lastname: Lee
mesh_terms: D001407:Bacillus; D001426:Bacterial Proteins / Q000737:chemistry / Q000187:drug effects*; D002352:Carrier Proteins / Q000737:chemistry / Q000187:drug effects*; D002414:Cations, Monovalent; D002942:Circular Dichroism; D009532:Nickel / Q000494:pharmacology*; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D012965:Sodium Chloride / Q000494:pharmacology*
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D002414:Cations, Monovalent; C090404:UreE protein, Bacteria; D012965:Sodium Chloride; C022838:nickel chloride; D009532:Nickel