Publikation:

Escherichia coli peptide binding protein OppA has a preference for positively charged peptides

Lade...
Vorschaubild

Dateien

Klepsch_2-4h7s8c8910dn2.PDF
Klepsch_2-4h7s8c8910dn2.PDFGröße: 920.23 KBDownloads: 29

Datum

2011

Autor:innen

Klepsch, Mirjam M.
Löw, Christian
Balbach, Jochen
Permentier, Hjalmar P.
Fusetti, Fabrizia
de Gier, Jan-Willem
Slotboom, Dirk J.
Berntsson, Ronnie Per Arne

Herausgeber:innen

Kontakt

ISSN der Zeitschrift

Electronic ISSN

ISBN

Bibliografische Daten

Verlag

Schriftenreihe

Auflagebezeichnung

ArXiv-ID

Internationale Patentnummer

Link zur Lizenz
oops

Angaben zur Forschungsförderung

Projekt

Open Access-Veröffentlichung
Open Access Green
Core Facility der Universität Konstanz

Gesperrt bis

Titel in einer weiteren Sprache

Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published

Erschienen in

Journal of molecular biology. 2011, 414(1), pp. 75-85. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2011.09.043

Zusammenfassung

The Escherichia coli peptide binding protein OppA is an essential component of the oligopeptide transporter Opp. Based on studies on its orthologue from Salmonella typhimurium, it has been proposed that OppA binds peptides between two and five amino acids long, with no apparent sequence selectivity. Here, we studied peptide binding to E. coli OppA directly and show that the protein has an unexpected preference for basic peptides. OppA was expressed in the periplasm, where it bound to available peptides. The protein was purified in complex with tightly bound peptides. The crystal structure (up to 2.0 Å) of OppA liganded with the peptides indicated that the protein has a preference for peptides containing a lysine. Mass spectrometry analysis of the bound peptides showed that peptides between two and five amino acids long bind to the protein and indeed hinted at a preference for positively charged peptides. The preference of OppA for peptides with basic residues, in particular lysines, was corroborated by binding studies with peptides of defined sequence using isothermal titration calorimetry and intrinsic protein fluorescence titration. The protein bound tripeptides and tetrapeptides containing positively charged residues with high affinity, whereas related peptides without lysines/arginines were bound with low affinity. A structure of OppA in an open conformation in the absence of ligands was also determined to 2.0 Å, revealing that the initial binding site displays a negative surface charge, consistent with the observed preference for positively charged peptides. Taken together, E. coli OppA appears to have a preference for basic peptides.

Zusammenfassung in einer weiteren Sprache

Fachgebiet (DDC)
540 Chemie

Schlagwörter

peptide binding protein, oligopeptide transporter, ABC transporter, Escherichia coli, substrate binding protein

Konferenz

Rezension
undefined / . - undefined, undefined

Forschungsvorhaben

Organisationseinheiten

Zeitschriftenheft

Zugehörige Datensätze in KOPS

Zitieren

ISO 690KLEPSCH, Mirjam M., Michael KOVERMANN, Christian LÖW, Jochen BALBACH, Hjalmar P. PERMENTIER, Fabrizia FUSETTI, Jan-Willem DE GIER, Dirk J. SLOTBOOM, Ronnie Per Arne BERNTSSON, 2011. Escherichia coli peptide binding protein OppA has a preference for positively charged peptides. In: Journal of molecular biology. 2011, 414(1), pp. 75-85. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2011.09.043
BibTex
@article{Klepsch2011-11-18Esche-44434,
  year={2011},
  doi={10.1016/j.jmb.2011.09.043},
  title={Escherichia coli peptide binding protein OppA has a preference for positively charged peptides},
  number={1},
  volume={414},
  issn={0022-2836},
  journal={Journal of molecular biology},
  pages={75--85},
  author={Klepsch, Mirjam M. and Kovermann, Michael and Löw, Christian and Balbach, Jochen and Permentier, Hjalmar P. and Fusetti, Fabrizia and de Gier, Jan-Willem and Slotboom, Dirk J. and Berntsson, Ronnie Per Arne}
}
RDF
<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/44434">
    <dcterms:title>Escherichia coli peptide binding protein OppA has a preference for positively charged peptides</dcterms:title>
    <dc:contributor>Balbach, Jochen</dc:contributor>
    <dc:creator>Berntsson, Ronnie Per Arne</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-01-08T09:00:07Z</dc:date>
    <dc:language>eng</dc:language>
    <dc:contributor>Permentier, Hjalmar P.</dc:contributor>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:contributor>Löw, Christian</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-01-08T09:00:07Z</dcterms:available>
    <dcterms:issued>2011-11-18</dcterms:issued>
    <dc:contributor>Klepsch, Mirjam M.</dc:contributor>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dc:contributor>Fusetti, Fabrizia</dc:contributor>
    <dc:contributor>Kovermann, Michael</dc:contributor>
    <dc:creator>Löw, Christian</dc:creator>
    <dc:creator>Balbach, Jochen</dc:creator>
    <dcterms:abstract xml:lang="eng">The Escherichia coli peptide binding protein OppA is an essential component of the oligopeptide transporter Opp. Based on studies on its orthologue from Salmonella typhimurium, it has been proposed that OppA binds peptides between two and five amino acids long, with no apparent sequence selectivity. Here, we studied peptide binding to E. coli OppA directly and show that the protein has an unexpected preference for basic peptides. OppA was expressed in the periplasm, where it bound to available peptides. The protein was purified in complex with tightly bound peptides. The crystal structure (up to 2.0 Å) of OppA liganded with the peptides indicated that the protein has a preference for peptides containing a lysine. Mass spectrometry analysis of the bound peptides showed that peptides between two and five amino acids long bind to the protein and indeed hinted at a preference for positively charged peptides. The preference of OppA for peptides with basic residues, in particular lysines, was corroborated by binding studies with peptides of defined sequence using isothermal titration calorimetry and intrinsic protein fluorescence titration. The protein bound tripeptides and tetrapeptides containing positively charged residues with high affinity, whereas related peptides without lysines/arginines were bound with low affinity. A structure of OppA in an open conformation in the absence of ligands was also determined to 2.0 Å, revealing that the initial binding site displays a negative surface charge, consistent with the observed preference for positively charged peptides. Taken together, E. coli OppA appears to have a preference for basic peptides.</dcterms:abstract>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/44434"/>
    <dc:creator>Klepsch, Mirjam M.</dc:creator>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/44434/1/Klepsch_2-4h7s8c8910dn2.PDF"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/44434/1/Klepsch_2-4h7s8c8910dn2.PDF"/>
    <dc:contributor>de Gier, Jan-Willem</dc:contributor>
    <dc:creator>Fusetti, Fabrizia</dc:creator>
    <dc:creator>de Gier, Jan-Willem</dc:creator>
    <dc:contributor>Berntsson, Ronnie Per Arne</dc:contributor>
    <dc:creator>Permentier, Hjalmar P.</dc:creator>
    <dc:creator>Kovermann, Michael</dc:creator>
    <dc:contributor>Slotboom, Dirk J.</dc:contributor>
    <dc:creator>Slotboom, Dirk J.</dc:creator>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
  </rdf:Description>
</rdf:RDF>

Interner Vermerk

xmlui.Submission.submit.DescribeStep.inputForms.label.kops_note_fromSubmitter

Kontakt
URL der Originalveröffentl.

Prüfdatum der URL

Prüfungsdatum der Dissertation

Finanzierungsart

Kommentar zur Publikation

Allianzlizenz
Corresponding Authors der Uni Konstanz vorhanden
Internationale Co-Autor:innen
Universitätsbibliographie
Nein
Begutachtet
Ja
Diese Publikation teilen