Publikation:

Binding of gephyrin to microtubules is regulated by its phosphorylation at Ser270

Lade...
Vorschaubild

Dateien

Zhou_2-1x1vpx4z3db987.pdf
Zhou_2-1x1vpx4z3db987.pdfGröße: 1.24 MBDownloads: 83

Datum

2021

Autor:innen

Zhou, Lin
Kiss, Éva
Kirsch, Joachim
Nawrotzki, Ralph Alexander
Kuhse, Jochen

Herausgeber:innen

Kontakt

ISSN der Zeitschrift

Electronic ISSN

ISBN

Bibliografische Daten

Verlag

Schriftenreihe

Auflagebezeichnung

ArXiv-ID

Internationale Patentnummer

Link zur Lizenz

Angaben zur Forschungsförderung

Projekt

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

Gesperrt bis

Titel in einer weiteren Sprache

Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published

Erschienen in

Histochemistry and Cell Biology. Springer. 2021, 156(1), pp. 5-18. ISSN 0948-6143. eISSN 1432-119X. Available under: doi: 10.1007/s00418-021-01973-2

Zusammenfassung

Gephyrin is a multifunctional scaffolding protein anchoring glycine- and subtypes of GABA type A- receptors at inhibitory postsynaptic membrane specializations by binding to the microtubule (MT) and/or the actin cytoskeleton. However, the conditions under which gephyrin can bind to MTs and its regulation are currently unknown. Here, we demonstrate that during the purification of MTs from rat brain by sedimentation of polymerized tubulin using high-speed centrifugation a fraction of gephyrin was bound to MTs, whereas gephyrin phosphorylated at the CDK5-dependent site Ser270 was detached from MTs and remained in the soluble protein fraction. Moreover, after collybistin fostered phosphorylation at Ser270 the binding of a recombinant gephyrin to MTs was strongly reduced in co-sedimentation assays. Correspondingly, upon substitution of wild-type gephyrin with recombinant gephyrin carrying alanine mutations at putative CDK5 phosphorylation sites the binding of gephyrin to MTs was increased. Furthermore, the analysis of cultured HEK293T and U2OS cells by immunofluorescence-microscopy disclosed a dispersed and punctuated endogenous gephyrin immunoreactivity co-localizing with MTs which was evidently not phosphorylated at Ser270. Thus, our study provides additional evidence for the binding of gephyrin to MTs in brain tissue and in in vitro cell systems. More importantly, our findings indicate that gephyrin-MT binding is restricted to a specific gephyrin fraction and depicts phosphorylation of gephyrin as a regulatory mechanism of this process by showing that soluble gephyrin detached from MTs can be detected specifically with the mAb7a antibody, which recognizes the Ser270 phosphorylated- version of gephyrin.

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 690ZHOU, Lin, Éva KISS, Rebecca DEMMIG, Joachim KIRSCH, Ralph Alexander NAWROTZKI, Jochen KUHSE, 2021. Binding of gephyrin to microtubules is regulated by its phosphorylation at Ser270. In: Histochemistry and Cell Biology. Springer. 2021, 156(1), pp. 5-18. ISSN 0948-6143. eISSN 1432-119X. Available under: doi: 10.1007/s00418-021-01973-2
BibTex
@article{Zhou2021-07Bindi-53549,
  year={2021},
  doi={10.1007/s00418-021-01973-2},
  title={Binding of gephyrin to microtubules is regulated by its phosphorylation at Ser270},
  number={1},
  volume={156},
  issn={0948-6143},
  journal={Histochemistry and Cell Biology},
  pages={5--18},
  author={Zhou, Lin and Kiss, Éva and Demmig, Rebecca and Kirsch, Joachim and Nawrotzki, Ralph Alexander and Kuhse, Jochen}
}
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/123456789/53549">
    <dc:contributor>Demmig, Rebecca</dc:contributor>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:creator>Kuhse, Jochen</dc:creator>
    <dc:contributor>Zhou, Lin</dc:contributor>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2021-04-30T07:15:40Z</dc:date>
    <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by/4.0/"/>
    <dc:creator>Kiss, Éva</dc:creator>
    <dc:contributor>Kiss, Éva</dc:contributor>
    <dc:rights>Attribution 4.0 International</dc:rights>
    <dcterms:abstract xml:lang="eng">Gephyrin is a multifunctional scaffolding protein anchoring glycine- and subtypes of GABA type A- receptors at inhibitory postsynaptic membrane specializations by binding to the microtubule (MT) and/or the actin cytoskeleton. However, the conditions under which gephyrin can bind to MTs and its regulation are currently unknown. Here, we demonstrate that during the purification of MTs from rat brain by sedimentation of polymerized tubulin using high-speed centrifugation a fraction of gephyrin was bound to MTs, whereas gephyrin phosphorylated at the CDK5-dependent site Ser270 was detached from MTs and remained in the soluble protein fraction. Moreover, after collybistin fostered phosphorylation at Ser270 the binding of a recombinant gephyrin to MTs was strongly reduced in co-sedimentation assays. Correspondingly, upon substitution of wild-type gephyrin with recombinant gephyrin carrying alanine mutations at putative CDK5 phosphorylation sites the binding of gephyrin to MTs was increased. Furthermore, the analysis of cultured HEK293T and U2OS cells by immunofluorescence-microscopy disclosed a dispersed and punctuated endogenous gephyrin immunoreactivity co-localizing with MTs which was evidently not phosphorylated at Ser270. Thus, our study provides additional evidence for the binding of gephyrin to MTs in brain tissue and in in vitro cell systems. More importantly, our findings indicate that gephyrin-MT binding is restricted to a specific gephyrin fraction and depicts phosphorylation of gephyrin as a regulatory mechanism of this process by showing that soluble gephyrin detached from MTs can be detected specifically with the mAb7a antibody, which recognizes the Ser270 phosphorylated- version of gephyrin.</dcterms:abstract>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/53549/1/Zhou_2-1x1vpx4z3db987.pdf"/>
    <dc:contributor>Kirsch, Joachim</dc:contributor>
    <dc:creator>Nawrotzki, Ralph Alexander</dc:creator>
    <dc:contributor>Nawrotzki, Ralph Alexander</dc:contributor>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/53549"/>
    <dcterms:issued>2021-07</dcterms:issued>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/53549/1/Zhou_2-1x1vpx4z3db987.pdf"/>
    <dc:contributor>Kuhse, Jochen</dc:contributor>
    <dc:creator>Kirsch, Joachim</dc:creator>
    <dc:language>eng</dc:language>
    <dcterms:title>Binding of gephyrin to microtubules is regulated by its phosphorylation at Ser270</dcterms:title>
    <dc:creator>Zhou, Lin</dc:creator>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2021-04-30T07:15:40Z</dcterms:available>
    <dc:creator>Demmig, Rebecca</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
Ja
Begutachtet
Ja
Diese Publikation teilen