Publikation:

Binding specificity of Escherichia coli trigger factor

Lade...
Vorschaubild

Dateien

Datum

2001

Autor:innen

Patzelt, Holger
Rüdiger, Stefan
Brehmer, Dirk
Kramer, Günter
Vorderwülbecke, Sonja
Schaffitzel, Elke
Waitz, Andreas
Hesterkamp, Thomas
Dong, Liying
Schneider-Mergener, Jens

Herausgeber:innen

Kontakt

ISSN der Zeitschrift

Electronic ISSN

ISBN

Bibliografische Daten

Verlag

Schriftenreihe

Auflagebezeichnung

ArXiv-ID

Internationale Patentnummer

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

Proceedings of the National Academy of Sciences of the United States of America. 2001, 98(25), pp. 14244-14249. ISSN 0027-8424. eISSN 1091-6490. Available under: doi: 10.1073/pnas.261432298

Zusammenfassung

The ribosome-associated chaperone trigger factor (TF) assists the folding of newly synthesized cytosolic proteins in Escherichia coli. Here, we determined the substrate specificity of TF by examining its binding to 2842 membrane-coupled 13meric peptides. The binding motif of TF was identified as a stretch of eight amino acids, enriched in basic and aromatic residues and with a positive net charge. Fluorescence spectroscopy verified that TF exhibited a comparable substrate specificity for peptides in solution. The affinity to peptides in solution was low, indicating that TF requires ribosome association to create high local concentrations of nascent polypeptide substrates for productive interaction in vivo. Binding to membrane-coupled peptides occurred through the central peptidyl-prolyl-cis/trans isomerase (PPIase) domain of TF, however, independently of prolyl residues. Crosslinking experiments showed that a TF fragment containing the PPIase domain linked to the ribosome via the N-terminal domain is sufficient for interaction with nascent polypeptide substrates. Homology modeling of the PPIase domain revealed a conserved FKB (FK506-binding protein)- like binding pocket composed of exposed aromatic residues embedded in a groove with negative surface charge. The features of this groove complement well the determined substrate specificity of TF. Moreover, a mutation (E178V) in this putative substrate binding groove known to enhance PPIase activity also enhanced TF s association with a prolyl-free model peptide in solution and with nascent polypeptides. This result suggests that both prolylindependent binding of peptide substrates and peptidyl-prolyl isomerization involve the same binding site.

Zusammenfassung in einer weiteren Sprache

Fachgebiet (DDC)
570 Biowissenschaften, Biologie

Schlagwörter

Konferenz

Rezension
undefined / . - undefined, undefined

Forschungsvorhaben

Organisationseinheiten

Zeitschriftenheft

Zugehörige Datensätze in KOPS

Zitieren

ISO 690PATZELT, Holger, Stefan RÜDIGER, Dirk BREHMER, Günter KRAMER, Sonja VORDERWÜLBECKE, Elke SCHAFFITZEL, Andreas WAITZ, Thomas HESTERKAMP, Liying DONG, Jens SCHNEIDER-MERGENER, Bernd BUKAU, Elke DEUERLING, 2001. Binding specificity of Escherichia coli trigger factor. In: Proceedings of the National Academy of Sciences of the United States of America. 2001, 98(25), pp. 14244-14249. ISSN 0027-8424. eISSN 1091-6490. Available under: doi: 10.1073/pnas.261432298
BibTex
@article{Patzelt2001Bindi-7324,
  year={2001},
  doi={10.1073/pnas.261432298},
  title={Binding specificity of Escherichia coli trigger factor},
  number={25},
  volume={98},
  issn={0027-8424},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  pages={14244--14249},
  author={Patzelt, Holger and Rüdiger, Stefan and Brehmer, Dirk and Kramer, Günter and Vorderwülbecke, Sonja and Schaffitzel, Elke and Waitz, Andreas and Hesterkamp, Thomas and Dong, Liying and Schneider-Mergener, Jens and Bukau, Bernd and Deuerling, Elke}
}
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/123456789/7324">
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc-nd/2.0/"/>
    <dc:creator>Schaffitzel, Elke</dc:creator>
    <dcterms:issued>2001</dcterms:issued>
    <dc:contributor>Rüdiger, Stefan</dc:contributor>
    <dc:creator>Kramer, Günter</dc:creator>
    <dc:contributor>Patzelt, Holger</dc:contributor>
    <dcterms:abstract xml:lang="eng">The ribosome-associated chaperone trigger factor (TF) assists the folding of newly synthesized cytosolic proteins in Escherichia coli. Here, we determined the substrate specificity of TF by examining its binding to 2842 membrane-coupled 13meric peptides. The binding motif of TF was identified as a stretch of eight amino acids, enriched in basic and aromatic residues and with a positive net charge. Fluorescence spectroscopy verified that TF exhibited a comparable substrate specificity for peptides in solution. The affinity to peptides in solution was low, indicating that TF requires ribosome association to create high local concentrations of nascent polypeptide substrates for productive interaction in vivo. Binding to membrane-coupled peptides occurred through the central peptidyl-prolyl-cis/trans isomerase (PPIase) domain of TF, however, independently of prolyl residues. Crosslinking experiments showed that a TF fragment containing the PPIase domain linked to the ribosome via the N-terminal domain is sufficient for interaction with nascent polypeptide substrates. Homology modeling of the PPIase domain revealed a conserved FKB (FK506-binding protein)- like binding pocket composed of exposed aromatic residues embedded in a groove with negative surface charge. The features of this groove complement well the determined substrate specificity of TF. Moreover, a mutation (E178V) in this putative substrate binding groove known to enhance PPIase activity also enhanced TF s association with a prolyl-free model peptide in solution and with nascent polypeptides. This result suggests that both prolylindependent binding of peptide substrates and peptidyl-prolyl isomerization involve the same binding site.</dcterms:abstract>
    <dc:rights>Attribution-NonCommercial-NoDerivs 2.0 Generic</dc:rights>
    <dc:creator>Vorderwülbecke, Sonja</dc:creator>
    <dc:creator>Hesterkamp, Thomas</dc:creator>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7324/1/Binding_specificity_of_Escherichia_coli_trigger_factor.pdf"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7324/1/Binding_specificity_of_Escherichia_coli_trigger_factor.pdf"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:33:32Z</dcterms:available>
    <dc:creator>Rüdiger, Stefan</dc:creator>
    <dc:contributor>Schneider-Mergener, Jens</dc:contributor>
    <dc:contributor>Brehmer, Dirk</dc:contributor>
    <dc:contributor>Bukau, Bernd</dc:contributor>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dc:contributor>Waitz, Andreas</dc:contributor>
    <dc:creator>Schneider-Mergener, Jens</dc:creator>
    <dc:creator>Dong, Liying</dc:creator>
    <dc:contributor>Kramer, Günter</dc:contributor>
    <dc:contributor>Deuerling, Elke</dc:contributor>
    <dc:contributor>Dong, Liying</dc:contributor>
    <dc:creator>Deuerling, Elke</dc:creator>
    <dc:language>eng</dc:language>
    <dc:creator>Waitz, Andreas</dc:creator>
    <dcterms:title>Binding specificity of Escherichia coli trigger factor</dcterms:title>
    <dc:contributor>Hesterkamp, Thomas</dc:contributor>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/7324"/>
    <dc:creator>Brehmer, Dirk</dc:creator>
    <dc:creator>Patzelt, Holger</dc:creator>
    <dcterms:bibliographicCitation>First publ. in: PNAS [Proceedings of the National Academy of Sciences], 98 (2004), pp. 14244-14249</dcterms:bibliographicCitation>
    <dc:contributor>Vorderwülbecke, Sonja</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:33:32Z</dc:date>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dc:format>application/pdf</dc:format>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:contributor>Schaffitzel, Elke</dc:contributor>
    <dc:creator>Bukau, Bernd</dc:creator>
  </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
Diese Publikation teilen