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Article: Structure, assembly, and topology of the G185R mutant of the fourth transmembrane domain of divalent metal transporter

TitleStructure, assembly, and topology of the G185R mutant of the fourth transmembrane domain of divalent metal transporter
Authors
Issue Date2005
PublisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/journals/jacsat/index.html
Citation
Journal Of The American Chemical Society, 2005, v. 127 n. 5, p. 1414-1423 How to Cite?
AbstractThe mammalian iron transporter, divalent metal transporter (DMT1), is a 12-transmembrane domain integral protein, responsible for dietary iron uptake in the duodenum and iron acquisition from transferrin in peripheral tissues. Two disease-causing mutants in animals have been found and attributed to the same missense mutation (G185R), which occurs within the putative transmembrane domain 4 (TM4) of DMT1. We have characterized a synthetic 24-mer peptide, corresponding to the sequence of the TM4 of DMT1 with G185R mutation using circular dichroism (CD) and NMR spectroscopy and show that the G185R peptide assumes mainly α-helical conformations in various membrane-mimetic environments. Solution structures derived from NMR and molecular dynamics/simulated annealing calculations demonstrate that the peptide exhibits a highly defined α-helix in its middle portion, flanked by a highly flexible N-terminus and a relatively ordered C-terminus. Both the folding and location of the C-terminus in SDS micelles are regulated by pH values. Paramagnetic broadening on peptide NMR signals by spin-labeled 5- and 16-doxylstearic acids and Mn 2+ ion suggests that both the N-terminus and the helical region of the peptide are embedded in SDS micelles. Surprisingly, self-association of the peptides for both the wild type and the G185R mutant studied by CD, electrospray ionization mass spectrometry, and NMR diffusion-ordered spectroscopy demonstrated that mutation of the Gly185 to a bulky and positively charged arginine causes a different self-assembly of the peptide, e.g., from a trimer to a hexamer, which implies that the quaternary structure of integral DMT1 may be crucial for its function in vivo.
Persistent Identifierhttp://hdl.handle.net/10722/167892
ISSN
2015 Impact Factor: 13.038
2015 SCImago Journal Rankings: 7.123
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorLi, Fen_US
dc.contributor.authorLi, Hen_US
dc.contributor.authorHu, Len_US
dc.contributor.authorKwan, Men_US
dc.contributor.authorChen, Gen_US
dc.contributor.authorHe, QYen_US
dc.contributor.authorSun, Hen_US
dc.date.accessioned2012-10-08T03:12:36Z-
dc.date.available2012-10-08T03:12:36Z-
dc.date.issued2005en_US
dc.identifier.citationJournal Of The American Chemical Society, 2005, v. 127 n. 5, p. 1414-1423en_US
dc.identifier.issn0002-7863en_US
dc.identifier.urihttp://hdl.handle.net/10722/167892-
dc.description.abstractThe mammalian iron transporter, divalent metal transporter (DMT1), is a 12-transmembrane domain integral protein, responsible for dietary iron uptake in the duodenum and iron acquisition from transferrin in peripheral tissues. Two disease-causing mutants in animals have been found and attributed to the same missense mutation (G185R), which occurs within the putative transmembrane domain 4 (TM4) of DMT1. We have characterized a synthetic 24-mer peptide, corresponding to the sequence of the TM4 of DMT1 with G185R mutation using circular dichroism (CD) and NMR spectroscopy and show that the G185R peptide assumes mainly α-helical conformations in various membrane-mimetic environments. Solution structures derived from NMR and molecular dynamics/simulated annealing calculations demonstrate that the peptide exhibits a highly defined α-helix in its middle portion, flanked by a highly flexible N-terminus and a relatively ordered C-terminus. Both the folding and location of the C-terminus in SDS micelles are regulated by pH values. Paramagnetic broadening on peptide NMR signals by spin-labeled 5- and 16-doxylstearic acids and Mn 2+ ion suggests that both the N-terminus and the helical region of the peptide are embedded in SDS micelles. Surprisingly, self-association of the peptides for both the wild type and the G185R mutant studied by CD, electrospray ionization mass spectrometry, and NMR diffusion-ordered spectroscopy demonstrated that mutation of the Gly185 to a bulky and positively charged arginine causes a different self-assembly of the peptide, e.g., from a trimer to a hexamer, which implies that the quaternary structure of integral DMT1 may be crucial for its function in vivo.en_US
dc.languageengen_US
dc.publisherAmerican Chemical Society. The Journal's web site is located at http://pubs.acs.org/journals/jacsat/index.htmlen_US
dc.relation.ispartofJournal of the American Chemical Societyen_US
dc.subject.meshAmino Acid Sequenceen_US
dc.subject.meshAnimalsen_US
dc.subject.meshCation Transport Proteins - Chemistry - Genetics - Metabolismen_US
dc.subject.meshCell Membrane - Chemistry - Metabolismen_US
dc.subject.meshCircular Dichroismen_US
dc.subject.meshIron-Binding Proteins - Chemistry - Genetics - Metabolismen_US
dc.subject.meshManganese - Chemistryen_US
dc.subject.meshMicellesen_US
dc.subject.meshModels, Molecularen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshMutation, Missenseen_US
dc.subject.meshNuclear Magnetic Resonance, Biomolecularen_US
dc.subject.meshProtein Structure, Secondaryen_US
dc.subject.meshProtein Structure, Tertiaryen_US
dc.subject.meshRatsen_US
dc.subject.meshSodium Dodecyl Sulfate - Chemistryen_US
dc.subject.meshSolventsen_US
dc.subject.meshSpectrometry, Mass, Electrospray Ionizationen_US
dc.subject.meshSpin Labelsen_US
dc.subject.meshTemperatureen_US
dc.titleStructure, assembly, and topology of the G185R mutant of the fourth transmembrane domain of divalent metal transporteren_US
dc.typeArticleen_US
dc.identifier.emailChen, G:ghc@yangtze.hku.hken_US
dc.identifier.emailSun, H:hsun@hkucc.hku.hken_US
dc.identifier.authorityChen, G=rp00671en_US
dc.identifier.authoritySun, H=rp00777en_US
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1021/ja047148ten_US
dc.identifier.pmid15686373-
dc.identifier.scopuseid_2-s2.0-13444292949en_US
dc.identifier.hkuros121038-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-13444292949&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume127en_US
dc.identifier.issue5en_US
dc.identifier.spage1414en_US
dc.identifier.epage1423en_US
dc.identifier.isiWOS:000226843900033-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridLi, F=36079222200en_US
dc.identifier.scopusauthoridLi, H=14023043100en_US
dc.identifier.scopusauthoridHu, L=7401557295en_US
dc.identifier.scopusauthoridKwan, M=35187349100en_US
dc.identifier.scopusauthoridChen, G=35253368600en_US
dc.identifier.scopusauthoridHe, QY=34770287900en_US
dc.identifier.scopusauthoridSun, H=7404827446en_US
dc.identifier.citeulike3813853-

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