Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET

Lade...
Vorschaubild
Dateien
Kahra_2-gl1wfvq8hbc87.PDF
Kahra_2-gl1wfvq8hbc87.PDFGröße: 373.05 KBDownloads: 8
Datum
2011
Autor:innen
Kahra, Dana
Löw, Christian
Hirschfeld, Verena
Balbach, Jochen
Hübner, Christian Gerhard
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
Sammlungen
Core Facility der Universität Konstanz
Gesperrt bis
Titel in einer weiteren Sprache
Forschungsvorhaben
Organisationseinheiten
Zeitschriftenheft
Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published
Erschienen in
Zusammenfassung

The relation between conformational dynamics and chemistry in enzyme catalysis recently has received increasing attention. While, in the past, the mechanochemical coupling was mainly attributed to molecular motors, nowadays, it seems that this linkage is far more general. Single-molecule fluorescence methods are perfectly suited to directly evidence conformational flexibility and dynamics. By labeling the enzyme SlyD, a member of peptidyl-prolyl cis-trans isomerases of the FK506 binding protein type with an inserted chaperone domain, with donor and acceptor fluorophores for single-molecule fluorescence resonance energy transfer, we directly monitor conformational flexibility and conformational dynamics between the chaperone domain and the FK506 binding protein domain. We find a broad distribution of distances between the labels with two main maxima, which we attribute to an open conformation and to a closed conformation of the enzyme. Correlation analysis demonstrates that the conformations exchange on a rate in the 100 Hz range. With the aid from Monte Carlo simulations, we show that there must be conformational flexibility beyond the two main conformational states. Interestingly, neither the conformational distribution nor the dynamics is significantly altered upon binding of substrates or other known binding partners. Based on these experimental findings, we propose a model where the conformational dynamics is used to search the conformation enabling the chemical step, which also explains the remarkable substrate promiscuity connected with a high efficiency of this class of peptidyl-prolyl cis-trans isomerases.

Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
540 Chemie
Schlagwörter
conformational dynamics; enzyme catalysis; protein folding; single molecule
Konferenz
Rezension
undefined / . - undefined, undefined
Zitieren
ISO 690KAHRA, Dana, Michael KOVERMANN, Christian LÖW, Verena HIRSCHFELD, Caroline HAUPT, Jochen BALBACH, Christian Gerhard HÜBNER, 2011. Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET. In: Journal of Molecular Biology (JMB). 2011, 411(4), pp. 781-790. ISSN 0022-2836. eISSN 1089-8638. Available under: doi: 10.1016/j.jmb.2011.05.002
BibTex
@article{Kahra2011-08-26Confo-44626,
  year={2011},
  doi={10.1016/j.jmb.2011.05.002},
  title={Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET},
  number={4},
  volume={411},
  issn={0022-2836},
  journal={Journal of Molecular Biology (JMB)},
  pages={781--790},
  author={Kahra, Dana and Kovermann, Michael and Löw, Christian and Hirschfeld, Verena and Haupt, Caroline and Balbach, Jochen and Hübner, Christian Gerhard}
}
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/123456789/44626">
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/44626/1/Kahra_2-gl1wfvq8hbc87.PDF"/>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-01-18T13:40:28Z</dc:date>
    <dc:contributor>Haupt, Caroline</dc:contributor>
    <dc:creator>Löw, Christian</dc:creator>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-01-18T13:40:28Z</dcterms:available>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dcterms:title>Conformational Plasticity and Dynamics in the Generic Protein Folding Catalyst SlyD Unraveled by Single-Molecule FRET</dcterms:title>
    <dc:creator>Hirschfeld, Verena</dc:creator>
    <dc:creator>Kahra, Dana</dc:creator>
    <dc:contributor>Hübner, Christian Gerhard</dc:contributor>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dc:contributor>Hirschfeld, Verena</dc:contributor>
    <dc:creator>Kovermann, Michael</dc:creator>
    <dc:contributor>Balbach, Jochen</dc:contributor>
    <dc:contributor>Löw, Christian</dc:contributor>
    <dc:creator>Balbach, Jochen</dc:creator>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/44626/1/Kahra_2-gl1wfvq8hbc87.PDF"/>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/44626"/>
    <dc:language>eng</dc:language>
    <dc:creator>Hübner, Christian Gerhard</dc:creator>
    <dc:contributor>Kahra, Dana</dc:contributor>
    <dc:creator>Haupt, Caroline</dc:creator>
    <dc:contributor>Kovermann, Michael</dc:contributor>
    <dcterms:abstract xml:lang="eng">The relation between conformational dynamics and chemistry in enzyme catalysis recently has received increasing attention. While, in the past, the mechanochemical coupling was mainly attributed to molecular motors, nowadays, it seems that this linkage is far more general. Single-molecule fluorescence methods are perfectly suited to directly evidence conformational flexibility and dynamics. By labeling the enzyme SlyD, a member of peptidyl-prolyl cis-trans isomerases of the FK506 binding protein type with an inserted chaperone domain, with donor and acceptor fluorophores for single-molecule fluorescence resonance energy transfer, we directly monitor conformational flexibility and conformational dynamics between the chaperone domain and the FK506 binding protein domain. We find a broad distribution of distances between the labels with two main maxima, which we attribute to an open conformation and to a closed conformation of the enzyme. Correlation analysis demonstrates that the conformations exchange on a rate in the 100 Hz range. With the aid from Monte Carlo simulations, we show that there must be conformational flexibility beyond the two main conformational states. Interestingly, neither the conformational distribution nor the dynamics is significantly altered upon binding of substrates or other known binding partners. Based on these experimental findings, we propose a model where the conformational dynamics is used to search the conformation enabling the chemical step, which also explains the remarkable substrate promiscuity connected with a high efficiency of this class of peptidyl-prolyl cis-trans isomerases.</dcterms:abstract>
    <dcterms:issued>2011-08-26</dcterms:issued>
  </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