Publikation:

Identification of novel functional mini-receptors by combinatorial screening of split-WW domains

Lade...
Vorschaubild

Dateien

Neitz_2-1acae5s7u8ycx9.pdf
Neitz_2-1acae5s7u8ycx9.pdfGröße: 1.53 MBDownloads: 19

Datum

2022

Autor:innen

Neitz, Hermann
Paul, Niels Benjamin
Häge, Florian R.
Lindner, Christina
Graebner, Roman
Thomas, Franziska

Herausgeber:innen

Kontakt

ISSN der Zeitschrift

Electronic ISSN

ISBN

Bibliografische Daten

Verlag

Schriftenreihe

Auflagebezeichnung

DOI (zitierfähiger Link)
ArXiv-ID

Internationale Patentnummer

Angaben zur Forschungsförderung

Projekt

Open Access-Veröffentlichung
Open Access Hybrid
Core Facility der Universität Konstanz
NMR Core Facility

Gesperrt bis

Titel in einer weiteren Sprache

Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published

Erschienen in

Chemical Science. Royal Society of Chemistry (RSC). 2022, 13(31), S. 9079-9090. ISSN 2041-6520. eISSN 2041-6539. Verfügbar unter: doi: 10.1039/d2sc01078j

Zusammenfassung

β-Sheet motifs such as the WW domain are increasingly being explored as building blocks for synthetic biological applications. Since the sequence-structure relationships of β-sheet motifs are generally complex compared to the well-studied α-helical coiled coil (CC), other approaches such as combinatorial screening should be included to vary the function of the peptide. In this study, we present a combinatorial approach to identify novel functional mini-proteins based on the WW-domain scaffold, which takes advantage of the successful reconstitution of the fragmented WW domain of hPin1 (hPin1WW) by CC association. Fragmentation of hPin1WW was performed in both loop 1 (CC-hPin1WW-L1) and loop 2 (CC-hPin1WW-L2), and the respective fragments were linked to the strands of an antiparallel heterodimeric CC. Structural analysis by CD and NMR spectroscopy revealed structural reconstitution of the WW-domain scaffold only in CC-hPin1WW-L1, but not in CC-hPin1WW-L2. Furthermore, by using 1H–15N HSQC NMR, fluorescence and CD spectroscopy, we demonstrated that binding properties of fragmented hPin1WW in CC-hPin1WW-L1 were fully restored by CC association. To demonstrate the power of this approach as a combinatorial screening platform, we synthesized a four-by-six library of N- and C-terminal hPin1WW-CC peptide fragments that was screened for a WW domain that preferentially binds to ATP over cAMP, phophocholine, or IP6. Using this screening platform, we identified one WW domain, which specifically binds ATP, and a phosphorylcholine-specific WW-based mini-receptor, both having binding dissociation constants in the lower micromolar range.

Zusammenfassung in einer weiteren Sprache

Fachgebiet (DDC)
540 Chemie

Schlagwörter

Konferenz

Rezension
undefined / . - undefined, undefined

Forschungsvorhaben

Organisationseinheiten

Zeitschriftenheft

Zugehörige Datensätze in KOPS

Zitieren

ISO 690NEITZ, Hermann, Niels Benjamin PAUL, Florian R. HÄGE, Christina LINDNER, Roman GRAEBNER, Michael KOVERMANN, Franziska THOMAS, 2022. Identification of novel functional mini-receptors by combinatorial screening of split-WW domains. In: Chemical Science. Royal Society of Chemistry (RSC). 2022, 13(31), S. 9079-9090. ISSN 2041-6520. eISSN 2041-6539. Verfügbar unter: doi: 10.1039/d2sc01078j
BibTex
@article{Neitz2022-08-10Ident-58235,
  year={2022},
  doi={10.1039/d2sc01078j},
  title={Identification of novel functional mini-receptors by combinatorial screening of split-WW domains},
  number={31},
  volume={13},
  issn={2041-6520},
  journal={Chemical Science},
  pages={9079--9090},
  author={Neitz, Hermann and Paul, Niels Benjamin and Häge, Florian R. and Lindner, Christina and Graebner, Roman and Kovermann, Michael and Thomas, Franziska}
}
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/123456789/58235">
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dc:creator>Lindner, Christina</dc:creator>
    <dc:contributor>Häge, Florian R.</dc:contributor>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/58235/1/Neitz_2-1acae5s7u8ycx9.pdf"/>
    <dc:language>eng</dc:language>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/58235"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/58235/1/Neitz_2-1acae5s7u8ycx9.pdf"/>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:contributor>Paul, Niels Benjamin</dc:contributor>
    <dc:rights>Attribution-NonCommercial 3.0 Unported</dc:rights>
    <dcterms:issued>2022-08-10</dcterms:issued>
    <dc:contributor>Lindner, Christina</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-08-04T13:09:33Z</dcterms:available>
    <dc:creator>Kovermann, Michael</dc:creator>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:contributor>Kovermann, Michael</dc:contributor>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dc:contributor>Thomas, Franziska</dc:contributor>
    <dc:creator>Paul, Niels Benjamin</dc:creator>
    <dc:creator>Graebner, Roman</dc:creator>
    <dc:contributor>Neitz, Hermann</dc:contributor>
    <dc:creator>Häge, Florian R.</dc:creator>
    <dcterms:title>Identification of novel functional mini-receptors by combinatorial screening of split-WW domains</dcterms:title>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc/3.0/"/>
    <dc:creator>Neitz, Hermann</dc:creator>
    <dc:creator>Thomas, Franziska</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2022-08-04T13:09:33Z</dc:date>
    <dc:contributor>Graebner, Roman</dc:contributor>
    <dcterms:abstract xml:lang="eng">β-Sheet motifs such as the WW domain are increasingly being explored as building blocks for synthetic biological applications. Since the sequence-structure relationships of β-sheet motifs are generally complex compared to the well-studied α-helical coiled coil (CC), other approaches such as combinatorial screening should be included to vary the function of the peptide. In this study, we present a combinatorial approach to identify novel functional mini-proteins based on the WW-domain scaffold, which takes advantage of the successful reconstitution of the fragmented WW domain of hPin1 (hPin1WW) by CC association. Fragmentation of hPin1WW was performed in both loop 1 (CC-hPin1WW-L1) and loop 2 (CC-hPin1WW-L2), and the respective fragments were linked to the strands of an antiparallel heterodimeric CC. Structural analysis by CD and NMR spectroscopy revealed structural reconstitution of the WW-domain scaffold only in CC-hPin1WW-L1, but not in CC-hPin1WW-L2. Furthermore, by using 1H–15N HSQC NMR, fluorescence and CD spectroscopy, we demonstrated that binding properties of fragmented hPin1WW in CC-hPin1WW-L1 were fully restored by CC association. To demonstrate the power of this approach as a combinatorial screening platform, we synthesized a four-by-six library of N- and C-terminal hPin1WW-CC peptide fragments that was screened for a WW domain that preferentially binds to ATP over cAMP, phophocholine, or IP6. Using this screening platform, we identified one WW domain, which specifically binds ATP, and a phosphorylcholine-specific WW-based mini-receptor, both having binding dissociation constants in the lower micromolar range.</dcterms:abstract>
  </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
Ja
Diese Publikation teilen