Analysis of mammalian 20S proteasome biogenesis : the maturation of beta-subunits is an ordered two-step mechanism involving autocatalysis

Lade...
Vorschaubild
Dateien
Zu diesem Dokument gibt es keine Dateien.
Datum
1996
Autor:innen
Kraft, R.
Kostka, S.
Henklein, Petra
Frömmel, Cornelius
Löwe, J.
Huber, R.
Kloetzel, Peter-Michael
Schmidt, M.
Herausgeber:innen
Kontakt
ISSN der Zeitschrift
Electronic ISSN
ISBN
Bibliografische Daten
Verlag
Schriftenreihe
Auflagebezeichnung
URI (zitierfähiger Link)
DOI (zitierfähiger Link)
ArXiv-ID
Internationale Patentnummer
Angaben zur Forschungsförderung
Projekt
Open Access-Veröffentlichung
Sammlungen
Core Facility der Universität Konstanz
Gesperrt bis
Titel in einer weiteren Sprache
Forschungsvorhaben
Organisationseinheiten
Zeitschriftenheft
Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published
Erschienen in
The EMBO journal. 1996, 15(24), pp. 6887-6898. ISSN 0261-4189. eISSN 1460-2075
Zusammenfassung

Maturation of eukaryotic 20S proteasomes involves the processing of beta-subunits by limited proteolysis. To study the processing mechanism we analysed different point mutations of the beta-subunit LMP2 in transfected human T2 cells. Here we show that the presence of the intact Gly-1Thr1 consensus motif and Lys33 are essential for correct processing. Mutation of Thr1, the active site residue in mature subunits, or of Lys33, results in complete inhibition of processing at the consensus site. In addition, proprotein processing in vitro of wild-type LMP2, incorporated in immature 16S precursor complexes, can be blocked by a proteasome-specific inhibitor. While the processing of inhibitor-treated wild-type proprotein was completely prevented, the site-directed mutagenesis of LMP2 results in processing intermediates carrying an extension of 8-10 residues preceding Thr1, suggesting an additional cleavage event within the prosequence. Furthermore, exchange of mammalian prosequences interferes with processing efficiency and suggests subunit specificity. Based on our data we propose a model for self-activation of proteasomal beta-subunits in which residue Thr1 serves as nucleophile and Lys33 as proton donor/acceptor. We provide evidence that subunit processing of mammalian beta-subunits proceeds via a novel ordered two-step mechanism involving autocatalysis.

Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
570 Biowissenschaften, Biologie
Schlagwörter
Konferenz
Rezension
undefined / . - undefined, undefined
Zitieren
ISO 690SCHMIDTKE, Gunter, R. KRAFT, S. KOSTKA, Petra HENKLEIN, Cornelius FRÖMMEL, J. LÖWE, R. HUBER, Peter-Michael KLOETZEL, M. SCHMIDT, 1996. Analysis of mammalian 20S proteasome biogenesis : the maturation of beta-subunits is an ordered two-step mechanism involving autocatalysis. In: The EMBO journal. 1996, 15(24), pp. 6887-6898. ISSN 0261-4189. eISSN 1460-2075
BibTex
@article{Schmidtke1996-12-16Analy-38755,
  year={1996},
  title={Analysis of mammalian 20S proteasome biogenesis : the maturation of beta-subunits is an ordered two-step mechanism involving autocatalysis},
  number={24},
  volume={15},
  issn={0261-4189},
  journal={The EMBO journal},
  pages={6887--6898},
  author={Schmidtke, Gunter and Kraft, R. and Kostka, S. and Henklein, Petra and Frömmel, Cornelius and Löwe, J. and Huber, R. and Kloetzel, Peter-Michael and Schmidt, M.}
}
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/123456789/38755">
    <dc:creator>Schmidt, M.</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2017-05-05T15:21:31Z</dc:date>
    <dc:creator>Kloetzel, Peter-Michael</dc:creator>
    <dc:contributor>Kraft, R.</dc:contributor>
    <dcterms:abstract xml:lang="eng">Maturation of eukaryotic 20S proteasomes involves the processing of beta-subunits by limited proteolysis. To study the processing mechanism we analysed different point mutations of the beta-subunit LMP2 in transfected human T2 cells. Here we show that the presence of the intact Gly-1Thr1 consensus motif and Lys33 are essential for correct processing. Mutation of Thr1, the active site residue in mature subunits, or of Lys33, results in complete inhibition of processing at the consensus site. In addition, proprotein processing in vitro of wild-type LMP2, incorporated in immature 16S precursor complexes, can be blocked by a proteasome-specific inhibitor. While the processing of inhibitor-treated wild-type proprotein was completely prevented, the site-directed mutagenesis of LMP2 results in processing intermediates carrying an extension of 8-10 residues preceding Thr1, suggesting an additional cleavage event within the prosequence. Furthermore, exchange of mammalian prosequences interferes with processing efficiency and suggests subunit specificity. Based on our data we propose a model for self-activation of proteasomal beta-subunits in which residue Thr1 serves as nucleophile and Lys33 as proton donor/acceptor. We provide evidence that subunit processing of mammalian beta-subunits proceeds via a novel ordered two-step mechanism involving autocatalysis.</dcterms:abstract>
    <dc:contributor>Kloetzel, Peter-Michael</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2017-05-05T15:21:31Z</dcterms:available>
    <dcterms:issued>1996-12-16</dcterms:issued>
    <dc:creator>Kraft, R.</dc:creator>
    <dc:contributor>Schmidtke, Gunter</dc:contributor>
    <dc:contributor>Henklein, Petra</dc:contributor>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dcterms:title>Analysis of mammalian 20S proteasome biogenesis : the maturation of beta-subunits is an ordered two-step mechanism involving autocatalysis</dcterms:title>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dc:creator>Huber, R.</dc:creator>
    <dc:creator>Löwe, J.</dc:creator>
    <dc:contributor>Huber, R.</dc:contributor>
    <dc:creator>Frömmel, Cornelius</dc:creator>
    <dc:contributor>Löwe, J.</dc:contributor>
    <dc:creator>Schmidtke, Gunter</dc:creator>
    <dc:language>eng</dc:language>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:contributor>Frömmel, Cornelius</dc:contributor>
    <dc:contributor>Kostka, S.</dc:contributor>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
    <dc:creator>Kostka, S.</dc:creator>
    <dc:contributor>Schmidt, M.</dc:contributor>
    <dc:creator>Henklein, Petra</dc:creator>
    <bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/38755"/>
  </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