Quantitative analysis of the binding affinity of poly(ADP-ribose) to specific binding proteins as a function of chain length

Lade...
Vorschaubild
Datum
2007
Autor:innen
Herausgeber:innen
Kontakt
ISSN der Zeitschrift
eISSN
item.preview.dc.identifier.isbn
Bibliografische Daten
Verlag
Schriftenreihe
URI (zitierfähiger Link)
DOI (zitierfähiger Link)
ArXiv-ID
Internationale Patentnummer
EU-Projektnummer
Projekt
Open Access-Veröffentlichung
Sammlungen
Gesperrt bis
Titel in einer weiteren Sprache
Forschungsvorhaben
Organisationseinheiten
Zeitschriftenheft
Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Erschienen in
Nucleic Acids Research ; 35 (2007), 21. - e143. - ISSN 0305-1048. - eISSN 1362-4962
Zusammenfassung
Poly(ADP-ribose) (PAR) is synthesized by poly(ADP-ribose) polymerases in response to genotoxic stress and interacts non-covalently with DNA damage checkpoint and repair proteins. Here, we present a variety of techniques to analyze this interaction in terms of selectivity and affinity. In vitro synthesized PAR was end-labeled using a carbonyl-reactive biotin analog. Binding of HPLC-fractionated PAR chains to the tumor suppressor protein p53 and to the nucleotide excision repair protein XPA was assessed using a novel electrophoretic mobility shift assay (EMSA). Long ADP-ribose chains (55-mer) promoted the formation of three specific complexes with p53. Short PAR chains (16-mer) were also able to bind p53, yet forming only one defined complex. In contrast, XPA did not interact with short polymer, but produced a single complex with long PAR chains (55-mer). In addition, we performed surface plasmon resonance with immobilized PAR chains, which allowed establishing binding constants and confirmed the results obtained by EMSA. Taken together, we developed several new protocols permitting the quantitative characterization of PAR protein binding. Furthermore, we demonstrated that the affinity of the non-covalent PAR interactions with specific binding proteins (XPA, p53) can be very high (nanomolar range) and depends both on the PAR chain length and on the binding protein.
Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
540 Chemie
Schlagwörter
Konferenz
Rezension
undefined / . - undefined, undefined. - (undefined; undefined)
Zitieren
ISO 690FAHRER, Jörg, Ramon KRANASTER, Matthias ALTMEYER, Andreas MARX, Alexander BÜRKLE, 2007. Quantitative analysis of the binding affinity of poly(ADP-ribose) to specific binding proteins as a function of chain length. In: Nucleic Acids Research. 35(21), e143. ISSN 0305-1048. eISSN 1362-4962
BibTex
@article{Fahrer2007Quant-9548,
  year={2007},
  title={Quantitative analysis of the binding affinity of poly(ADP-ribose) to specific binding proteins as a function of chain length},
  number={21},
  volume={35},
  issn={0305-1048},
  journal={Nucleic Acids Research},
  author={Fahrer, Jörg and Kranaster, Ramon and Altmeyer, Matthias and Marx, Andreas and Bürkle, Alexander},
  note={Article Number: e143}
}
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/123456789/9548">
    <dcterms:abstract xml:lang="eng">Poly(ADP-ribose) (PAR) is synthesized by poly(ADP-ribose) polymerases in response to genotoxic stress and interacts non-covalently with DNA damage checkpoint and repair proteins. Here, we present a variety of techniques to analyze this interaction in terms of selectivity and affinity. In vitro synthesized PAR was end-labeled using a carbonyl-reactive biotin analog. Binding of HPLC-fractionated PAR chains to the tumor suppressor protein p53 and to the nucleotide excision repair protein XPA was assessed using a novel electrophoretic mobility shift assay (EMSA). Long ADP-ribose chains (55-mer) promoted the formation of three specific complexes with p53. Short PAR chains (16-mer) were also able to bind p53, yet forming only one defined complex. In contrast, XPA did not interact with short polymer, but produced a single complex with long PAR chains (55-mer). In addition, we performed surface plasmon resonance with immobilized PAR chains, which allowed establishing binding constants and confirmed the results obtained by EMSA. Taken together, we developed several new protocols permitting the quantitative characterization of PAR protein binding. Furthermore, we demonstrated that the affinity of the non-covalent PAR interactions with specific binding proteins (XPA, p53) can be very high (nanomolar range) and depends both on the PAR chain length and on the binding protein.</dcterms:abstract>
    <dc:contributor>Fahrer, Jörg</dc:contributor>
    <dc:creator>Fahrer, Jörg</dc:creator>
    <dc:creator>Kranaster, Ramon</dc:creator>
    <dcterms:bibliographicCitation>First publ. in: Nucleic Acids Research 35 (2007), 21, e143</dcterms:bibliographicCitation>
    <dc:language>eng</dc:language>
    <dc:rights>Attribution-NonCommercial-NoDerivs 2.0 Generic</dc:rights>
    <dc:contributor>Marx, Andreas</dc:contributor>
    <dc:creator>Marx, Andreas</dc:creator>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:contributor>Kranaster, Ramon</dc:contributor>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/9548"/>
    <dc:creator>Altmeyer, Matthias</dc:creator>
    <dc:contributor>Bürkle, Alexander</dc:contributor>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/9548/1/Fahrer_Nucl_Acids_Res_2007.pdf"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/9548/1/Fahrer_Nucl_Acids_Res_2007.pdf"/>
    <dc:format>application/pdf</dc:format>
    <dcterms:issued>2007</dcterms:issued>
    <dcterms:title>Quantitative analysis of the binding affinity of poly(ADP-ribose) to specific binding proteins as a function of chain length</dcterms:title>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T18:12:40Z</dcterms:available>
    <dc:creator>Bürkle, Alexander</dc:creator>
    <dc:contributor>Altmeyer, Matthias</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T18:12:40Z</dc:date>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc-nd/2.0/"/>
  </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
Ja
Begutachtet