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Article: Nickel Translocation between Metallochaperones HypA and UreE in Helicobacter pylori

TitleNickel Translocation between Metallochaperones HypA and UreE in Helicobacter pylori
Authors
Issue Date2014
PublisherThe Royal Society of Chemistry. The Journal's web site is located at http://www.rsc.org/Publishing/Journals/MT/About.asp
Citation
Metallomics, 2014, v. 6, p. 1731-1736 How to Cite?
AbstractIncorporation of nickel ions to the active sites of urease and hydrogenase is prerequisite for the appropriate functions of the metalloenzymes. Such a process requires the participation of several accessory proteins. Interestingly, some of them are shared by the two enzymes in their maturation processes. In this work, we characterized the molecular details of the interaction of metallochaperones UreE and HypA in Helicobacter pylori. We show by chemical cross-linking and static light scattering that the UreE dimer binds to HypA to form a hetero-complex i.e. HypA–(UreE)2. The dissociation constant (Kd) of the protein complex was determined by ITC to be 1 μM in the absence of nickel ions; whereas binding of Ni2+ but not Zn2+ to UreE resulted in ca. one fold decrease in the affinity. The putative interfaces on HypA unveiled by NMR chemical shift perturbation were found mainly at the nickel binding domain and in the cleft between α1 and β1/β6. We also identified that the C-domain of UreE, in particular the C-terminal residues of 158–170 are indispensable for the interaction of UreE and HypA. Such an interaction was also observed intracellularly by GFP-fragment reassembly assay. Moreover, we demonstrated using a fluorescent probe that nickel is transferred from HypA to UreE via the specific protein–protein interaction. Deletion of the C-terminus (residues 158–170) of UreE abolished nickel transfer and led to a significant decrease in urease activity. This study provides direct in vitro and in vivo evidence as well as molecular details of nickel translocation mediated by protein–protein interaction.
Persistent Identifierhttp://hdl.handle.net/10722/202448
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYANG, Xen_US
dc.contributor.authorLi, Hen_US
dc.contributor.authorCheng, Ten_US
dc.contributor.authorXia, Wen_US
dc.contributor.authorLai, AYTen_US
dc.contributor.authorSun, Hen_US
dc.date.accessioned2014-09-19T07:50:39Z-
dc.date.available2014-09-19T07:50:39Z-
dc.date.issued2014-
dc.identifier.citationMetallomics, 2014, v. 6, p. 1731-1736en_US
dc.identifier.urihttp://hdl.handle.net/10722/202448-
dc.description.abstractIncorporation of nickel ions to the active sites of urease and hydrogenase is prerequisite for the appropriate functions of the metalloenzymes. Such a process requires the participation of several accessory proteins. Interestingly, some of them are shared by the two enzymes in their maturation processes. In this work, we characterized the molecular details of the interaction of metallochaperones UreE and HypA in Helicobacter pylori. We show by chemical cross-linking and static light scattering that the UreE dimer binds to HypA to form a hetero-complex i.e. HypA–(UreE)2. The dissociation constant (Kd) of the protein complex was determined by ITC to be 1 μM in the absence of nickel ions; whereas binding of Ni2+ but not Zn2+ to UreE resulted in ca. one fold decrease in the affinity. The putative interfaces on HypA unveiled by NMR chemical shift perturbation were found mainly at the nickel binding domain and in the cleft between α1 and β1/β6. We also identified that the C-domain of UreE, in particular the C-terminal residues of 158–170 are indispensable for the interaction of UreE and HypA. Such an interaction was also observed intracellularly by GFP-fragment reassembly assay. Moreover, we demonstrated using a fluorescent probe that nickel is transferred from HypA to UreE via the specific protein–protein interaction. Deletion of the C-terminus (residues 158–170) of UreE abolished nickel transfer and led to a significant decrease in urease activity. This study provides direct in vitro and in vivo evidence as well as molecular details of nickel translocation mediated by protein–protein interaction.en_US
dc.languageengen_US
dc.publisherThe Royal Society of Chemistry. The Journal's web site is located at http://www.rsc.org/Publishing/Journals/MT/About.aspen_US
dc.relation.ispartofMetallomicsen_US
dc.titleNickel Translocation between Metallochaperones HypA and UreE in Helicobacter pylorien_US
dc.typeArticleen_US
dc.identifier.emailLi, H: hylichem@hku.hken_US
dc.identifier.emailCheng, T: chengtfc@hku.hken_US
dc.identifier.emailXia, W: weixia1984@hku.hken_US
dc.identifier.emailLai, AYT: skytc13@hku.hken_US
dc.identifier.emailSun, H: hsun@hku.hken_US
dc.identifier.authoritySun, H=rp00777en_US
dc.identifier.doi10.1039/C4MT00134Fen_US
dc.identifier.pmid25010720-
dc.identifier.scopuseid_2-s2.0-84906564755-
dc.identifier.hkuros235257en_US
dc.identifier.isiWOS:000341018500018-

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