Target-directed proteolysis at the ribosome
Dateien
Datum
Autor:innen
Herausgeber:innen
ISSN der Zeitschrift
Electronic ISSN
ISBN
Bibliografische Daten
Verlag
Schriftenreihe
Auflagebezeichnung
URI (zitierfähiger Link)
Internationale Patentnummer
Link zur Lizenz
Angaben zur Forschungsförderung
Projekt
Open Access-Veröffentlichung
Sammlungen
Core Facility der Universität Konstanz
Titel in einer weiteren Sprache
Publikationstyp
Publikationsstatus
Erschienen in
Zusammenfassung
Target directed proteolysis allows specific processing of proteins in vivo. This method uses tobacco etch virus (TEV) NIa protease that recognizes a seven-residue consensus sequence. Because of its specificity, proteins engineered to contain a cleavage site are proteolysed, whereas other proteins remain unaffected. Therefore, this approach can be used to study the structure and function of target proteins in their natural environment within living cells. One application is the conditional inactivation of essential proteins, which is based on the concept that a target containing a recognition site can be inactivated by coexpressed TEV protease. We have previously identified one site in the secretion factor SecA that tolerated a TEV protease site insert. Coexpression of TEV protease in the cytoplasm led to incomplete cleavage and a mild secretion defect. To improve the efficiency of proteolysis, TEV protease was attached to the ribosome. We show here that cleaving SecA under these conditions is one way of increasing the efficiency of target directed proteolysis. The implications of recruiting novel biological activities to ribosomes are discussed.
Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
Schlagwörter
Konferenz
Rezension
Zitieren
ISO 690
HENRICHS, Tanja, Natasha MIKHALEVA, Charlotte CONZ, Elke DEUERLING, Dana BOYD, Adrian ZELAZNY, Eitan BIBI, Nenad BAN, Michael EHRMANN, 2005. Target-directed proteolysis at the ribosome. In: PNAS [ Proceedings of the National Academy of Sciences ]. 2005, 102(12), pp. 4246-4251BibTex
@article{Henrichs2005Targe-7251, year={2005}, title={Target-directed proteolysis at the ribosome}, number={12}, volume={102}, journal={PNAS [ Proceedings of the National Academy of Sciences ]}, pages={4246--4251}, author={Henrichs, Tanja and Mikhaleva, Natasha and Conz, Charlotte and Deuerling, Elke and Boyd, Dana and Zelazny, Adrian and Bibi, Eitan and Ban, Nenad and Ehrmann, Michael} }
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/123456789/7251"> <dc:contributor>Boyd, Dana</dc:contributor> <dcterms:bibliographicCitation>First publ. in: PNAS [Proceedings of the National Academy of Sciences],102 (2005), 12, pp. 4246-4251</dcterms:bibliographicCitation> <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:32:58Z</dc:date> <dc:creator>Bibi, Eitan</dc:creator> <dc:rights>Attribution-NonCommercial-NoDerivs 2.0 Generic</dc:rights> <foaf:homepage rdf:resource="http://localhost:8080/"/> <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T17:32:58Z</dcterms:available> <dcterms:title>Target-directed proteolysis at the ribosome</dcterms:title> <dcterms:rights rdf:resource="http://creativecommons.org/licenses/by-nc-nd/2.0/"/> <dc:contributor>Henrichs, Tanja</dc:contributor> <dc:contributor>Deuerling, Elke</dc:contributor> <dc:creator>Zelazny, Adrian</dc:creator> <dc:creator>Ban, Nenad</dc:creator> <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/> <dc:creator>Conz, Charlotte</dc:creator> <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7251/1/Target_directed_proteolysis_at_the_ribosome.pdf"/> <dcterms:abstract xml:lang="eng">Target directed proteolysis allows specific processing of proteins in vivo. This method uses tobacco etch virus (TEV) NIa protease that recognizes a seven-residue consensus sequence. Because of its specificity, proteins engineered to contain a cleavage site are proteolysed, whereas other proteins remain unaffected. Therefore, this approach can be used to study the structure and function of target proteins in their natural environment within living cells. One application is the conditional inactivation of essential proteins, which is based on the concept that a target containing a recognition site can be inactivated by coexpressed TEV protease. We have previously identified one site in the secretion factor SecA that tolerated a TEV protease site insert. Coexpression of TEV protease in the cytoplasm led to incomplete cleavage and a mild secretion defect. To improve the efficiency of proteolysis, TEV protease was attached to the ribosome. We show here that cleaving SecA under these conditions is one way of increasing the efficiency of target directed proteolysis. The implications of recruiting novel biological activities to ribosomes are discussed.</dcterms:abstract> <dc:format>application/pdf</dc:format> <dc:contributor>Conz, Charlotte</dc:contributor> <dc:contributor>Zelazny, Adrian</dc:contributor> <dc:language>eng</dc:language> <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/7251/1/Target_directed_proteolysis_at_the_ribosome.pdf"/> <dc:creator>Ehrmann, Michael</dc:creator> <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/7251"/> <dc:contributor>Ehrmann, Michael</dc:contributor> <dc:creator>Deuerling, Elke</dc:creator> <dc:contributor>Mikhaleva, Natasha</dc:contributor> <dc:creator>Mikhaleva, Natasha</dc:creator> <dcterms:issued>2005</dcterms:issued> <dc:creator>Henrichs, Tanja</dc:creator> <dc:contributor>Ban, Nenad</dc:contributor> <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/> <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/> <dc:creator>Boyd, Dana</dc:creator> <dc:contributor>Bibi, Eitan</dc:contributor> </rdf:Description> </rdf:RDF>