File Download
  Links for fulltext
     (May Require Subscription)
Supplementary

Article: Metallo-GTPase HypB from helicobacter pylori and its interaction with nickel chaperone protein HypA

TitleMetallo-GTPase HypB from helicobacter pylori and its interaction with nickel chaperone protein HypA
Authors
Issue Date2012
PublisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/
Citation
Journal of Biological Chemistry, 2012, v. 287 n. 9, p. 6753-6763 How to Cite?
AbstractThe maturation of [NiFe]-hydrogenase is highly dependent on a battery of chaperone proteins. Among these, HypA and HypB were proposed to exert nickel delivery functions in the metallocenter assembly process, although the detailed mechanism remains unclear. Herein, we have overexpressed and purified wild-type HypB as well as two mutants, K168A and M186L/F190V, from Helicobacter pylori. We demonstrated that all proteins bind Ni(2+) at a stoichiometry of one Ni(2+) per monomer of the proteins with dissociation constants at micromolar levels. Ni(2+) elevated GTPase activity of WT HypB, which is attributable to a lower affinity of the protein toward GDP as well as Ni(2+)-induced dimerization. The disruption of GTP-dependent dimerization has led to GTPase activities of both mutants in apo-forms almost completely abolished, compared with the wild-type protein. The GTPase activity is partially restored for HypB(M186L/F190V) mutant but not for HypB(K168A) mutant upon Ni(2+) binding. HypB forms a complex with its partner protein HypA with a low affinity (K(d) of 52.2 +/- 8.8 muM). Such interactions were also observed in vivo both in the absence and presence of nickel using a GFP-fragment reassembly technique. The putative protein-protein interfaces on H. pylori HypA and HypB proteins were identified by NMR chemical shift perturbation and mutagenesis studies, respectively. Intriguingly, the unique N terminus of H. pylori HypB was identified to participate in the interaction with H. pylori HypA. These structural and functional studies provide insight into the molecular mechanism of Ni(2+) delivery during maturation of [NiFe]-hydrogenase.
Persistent Identifierhttp://hdl.handle.net/10722/168644
ISSN
2015 Impact Factor: 4.258
2015 SCImago Journal Rankings: 3.151
PubMed Central ID
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorXia, Wen_US
dc.contributor.authorLi, Hen_US
dc.contributor.authorYang, Xen_US
dc.contributor.authorWong, KBen_US
dc.contributor.authorSun, Hen_US
dc.date.accessioned2012-10-08T03:23:53Z-
dc.date.available2012-10-08T03:23:53Z-
dc.date.issued2012en_US
dc.identifier.citationJournal of Biological Chemistry, 2012, v. 287 n. 9, p. 6753-6763en_US
dc.identifier.issn0021-9258en_US
dc.identifier.urihttp://hdl.handle.net/10722/168644-
dc.description.abstractThe maturation of [NiFe]-hydrogenase is highly dependent on a battery of chaperone proteins. Among these, HypA and HypB were proposed to exert nickel delivery functions in the metallocenter assembly process, although the detailed mechanism remains unclear. Herein, we have overexpressed and purified wild-type HypB as well as two mutants, K168A and M186L/F190V, from Helicobacter pylori. We demonstrated that all proteins bind Ni(2+) at a stoichiometry of one Ni(2+) per monomer of the proteins with dissociation constants at micromolar levels. Ni(2+) elevated GTPase activity of WT HypB, which is attributable to a lower affinity of the protein toward GDP as well as Ni(2+)-induced dimerization. The disruption of GTP-dependent dimerization has led to GTPase activities of both mutants in apo-forms almost completely abolished, compared with the wild-type protein. The GTPase activity is partially restored for HypB(M186L/F190V) mutant but not for HypB(K168A) mutant upon Ni(2+) binding. HypB forms a complex with its partner protein HypA with a low affinity (K(d) of 52.2 +/- 8.8 muM). Such interactions were also observed in vivo both in the absence and presence of nickel using a GFP-fragment reassembly technique. The putative protein-protein interfaces on H. pylori HypA and HypB proteins were identified by NMR chemical shift perturbation and mutagenesis studies, respectively. Intriguingly, the unique N terminus of H. pylori HypB was identified to participate in the interaction with H. pylori HypA. These structural and functional studies provide insight into the molecular mechanism of Ni(2+) delivery during maturation of [NiFe]-hydrogenase.en_US
dc.languageengen_US
dc.publisherAmerican Society for Biochemistry and Molecular Biology, Inc. The Journal's web site is located at http://www.jbc.org/en_US
dc.relation.ispartofJournal of Biological Chemistryen_US
dc.rightsJournal of Biological Chemistry. Copyright © American Society for Biochemistry and Molecular Biology, Inc.-
dc.subject.meshBacterial Proteins - chemistry - genetics - metabolismen_US
dc.subject.meshGTP-Binding Proteins - chemistry - genetics - metabolismen_US
dc.subject.meshHelicobacter pylori - enzymology - geneticsen_US
dc.subject.meshMetallochaperones - genetics - metabolismen_US
dc.subject.meshNickel - metabolismen_US
dc.titleMetallo-GTPase HypB from helicobacter pylori and its interaction with nickel chaperone protein HypAen_US
dc.typeArticleen_US
dc.identifier.emailLi, H: hylichem@hku.hken_US
dc.identifier.emailSun, H: hsun@hku.hk-
dc.identifier.authoritySun, H=rp00777en_US
dc.description.naturelink_to_OA_fulltexten_US
dc.identifier.doi10.1074/jbc.M111.287581en_US
dc.identifier.pmid22179820-
dc.identifier.pmcidPMC3307304-
dc.identifier.scopuseid_2-s2.0-84863149329en_US
dc.identifier.hkuros205072-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-84863149329&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume287en_US
dc.identifier.issue9en_US
dc.identifier.spage6753en_US
dc.identifier.epage6763en_US
dc.identifier.isiWOS:000300791800061-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridSun, H=7404827446en_US
dc.identifier.scopusauthoridWong, KB=7404759301en_US
dc.identifier.scopusauthoridYang, X=55273211100en_US
dc.identifier.scopusauthoridLi, H=37063577200en_US
dc.identifier.scopusauthoridXia, W=40262950100en_US

Export via OAI-PMH Interface in XML Formats


OR


Export to Other Non-XML Formats