Please use this identifier to cite or link to this item: https://olympias.lib.uoi.gr/jspui/handle/123456789/23474
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dc.contributor.authorFrillingos, S.en
dc.contributor.authorWu, J.en
dc.contributor.authorVenkatesan, P.en
dc.contributor.authorKaback, H. R.en
dc.date.accessioned2015-11-24T19:32:56Z-
dc.date.available2015-11-24T19:32:56Z-
dc.identifier.issn0006-2960-
dc.identifier.urihttps://olympias.lib.uoi.gr/jspui/handle/123456789/23474-
dc.rightsDefault Licence-
dc.subjectAntibodies, Monoclonal/*chemistryen
dc.subject*Binding Sites, Antibody/geneticsen
dc.subjectCysteine/geneticsen
dc.subjectEscherichia coli/chemistry/*enzymologyen
dc.subject*Escherichia coli Proteinsen
dc.subjectLigandsen
dc.subjectMembrane Transport Proteins/*chemistry/genetics/*metabolismen
dc.subject*Monosaccharide Transport Proteinsen
dc.subjectMutagenesis, Site-Directeden
dc.subjectProtein Conformationen
dc.subject*Symportersen
dc.titleBinding of ligand or monoclonal antibody 4B1 induces discrete structural changes in the lactose permease of Escherichia colien
heal.typejournalArticle-
heal.type.enJournal articleen
heal.type.elΆρθρο Περιοδικούel
heal.identifier.primary10.1021/bi970233b-
heal.identifier.secondaryhttp://www.ncbi.nlm.nih.gov/pubmed/9174357-
heal.identifier.secondaryhttp://pubs.acs.org/doi/pdfplus/10.1021/bi970233b-
heal.languageen-
heal.accesscampus-
heal.recordProviderΠανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικήςel
heal.publicationDate1997-
heal.abstractBy using Cys-scanning mutagenesis with site-directed sulfhydryl modification in situ [Frillingos, S., & Kaback, H. R. (1996) Biochemistry 35, 3950-3956], conformational changes induced by binding of ligand or monoclonal antibody (mAb) 4B1 in the lactose permease of Escherichia coli were studied. Out of 31 single-Cys replacement mutants in helices I, V, VII, VIII, X, or XI, 4B1 binding alters the reactivity of Val238-->Cys (helix VII), Val331-->Cys (helix X), or single-Cys355 (helix XI) permease with N-ethylmaleimide (NEM) in right-side-out membrane vesicles. In addition, site-directed fluorescence spectroscopy shows that mAb 4B1 binding causes position 331 (helix X) in the permease to experience a more hydrophobic environment. In contrast, ligand binding elicits more widespread changes, as evidenced by enhancement of the NEM reactivity of Ala244-->Cys, Thr248-->Cys (helix VII), Thr265-->Cys (helix VIII), Val315-->Cys (helix X), Gln359-->Cys, or Met362-->Cys (helix XI) permease, none of which are altered by 4B1 binding. Furthermore, no effect of 4B1 is observed on the reactivity of Cys148 (helix V), Val264-->Cys, Gly268-->Cys, or Asn272-->Cys (helix VIII), positions which probably make direct contact with substrate. With respect to the N-terminal half of the permease, 4B1 binding causes a small increase in the reactivity of mutants Pro28-->Cys or Pro31-->Cys (helix I), while ligand binding causes much greater increases in reactivity. The findings indicate that 4B1 binding induces a structural change in the permease that is much less widespread than that induced by ligand binding.en
heal.journalNameBiochemistryen
heal.journalTypepeer-reviewed-
heal.fullTextAvailabilityTRUE-
Appears in Collections:Άρθρα σε επιστημονικά περιοδικά ( Ανοικτά) - ΙΑΤ

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