Publikation: Scalable, Non-denaturing Purification of Phosphoproteins Using Ga3+-IMAC : N2A and M1M2 Titin Components as Study case
Lade...
Dateien
Datum
2019
Autor:innen
Herausgeber:innen
ISSN der Zeitschrift
Electronic ISSN
ISBN
Bibliografische Daten
Verlag
Schriftenreihe
Auflagebezeichnung
URI (zitierfähiger Link)
DOI (zitierfähiger Link)
Internationale Patentnummer
Link zur Lizenz
Angaben zur Forschungsförderung
European Union (EU): 753054
Projekt
Development of novel computational biology pipeline for the efficient classification of titin SNPs for clinical use - TTNPred
Open Access-Veröffentlichung
Open Access Green
Sammlungen
Core Facility der Universität Konstanz
Titel in einer weiteren Sprache
Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published
Erschienen in
The Protein Journal. 2019, 38(2), pp. 181-189. ISSN 1572-3887. eISSN 1875-8355. Available under: doi: 10.1007/s10930-019-09815-w
Zusammenfassung
The purification of phosphorylated proteins in a folded state and in large enough quantity for biochemical or biophysical analysis remains a challenging task. Here, we develop a new implementation of the method of gallium immobilized metal chromatography (Ga3+-IMAC) as to permit the selective enrichment of phosphoproteins in the milligram scale and under native conditions using automated FPLC instrumentation. We apply this method to the purification of the UN2A and M1M2 components of the muscle protein titin upon being monophosphorylated in vitro by cAMP-dependent protein kinase (PKA). We found that UN2A is phosphorylated by PKA at its C-terminus in residue S9578 and M1M2 is phosphorylated in its interdomain linker sequence at position T32607. We demonstrate that the Ga3+-IMAC method is efficient, economical and suitable for implementation in automated purification pipelines for recombinant proteins. The procedure can be applied both to the selective enrichment and to the removal of phosphoproteins from biochemical samples.
Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
570 Biowissenschaften, Biologie
Schlagwörter
Konferenz
Rezension
undefined / . - undefined, undefined
Zitieren
ISO 690
ADAMS, Michael, Jennifer R. FLEMING, Eva RIEHLE, Tiankun ZHOU, Thomas ZACHARCHENKO, Marija MARKOVIC, Olga MAYANS, 2019. Scalable, Non-denaturing Purification of Phosphoproteins Using Ga3+-IMAC : N2A and M1M2 Titin Components as Study case. In: The Protein Journal. 2019, 38(2), pp. 181-189. ISSN 1572-3887. eISSN 1875-8355. Available under: doi: 10.1007/s10930-019-09815-wBibTex
@article{Adams2019-04Scala-45025,
year={2019},
doi={10.1007/s10930-019-09815-w},
title={Scalable, Non-denaturing Purification of Phosphoproteins Using Ga<sup>3+</sup>-IMAC : N2A and M1M2 Titin Components as Study case},
number={2},
volume={38},
issn={1572-3887},
journal={The Protein Journal},
pages={181--189},
author={Adams, Michael and Fleming, Jennifer R. and Riehle, Eva and Zhou, Tiankun and Zacharchenko, Thomas and Markovic, Marija and Mayans, Olga}
}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/123456789/45025">
<dc:contributor>Riehle, Eva</dc:contributor>
<void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
<dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
<dc:language>eng</dc:language>
<bibo:uri rdf:resource="https://kops.uni-konstanz.de/handle/123456789/45025"/>
<dc:contributor>Adams, Michael</dc:contributor>
<dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/>
<dc:creator>Riehle, Eva</dc:creator>
<dc:rights>terms-of-use</dc:rights>
<dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/45025/1/Adams_2-14disd67exh901.pdf"/>
<dc:creator>Markovic, Marija</dc:creator>
<dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-02-14T10:01:08Z</dcterms:available>
<dc:contributor>Zacharchenko, Thomas</dc:contributor>
<dc:creator>Adams, Michael</dc:creator>
<dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/28"/>
<dc:creator>Zhou, Tiankun</dc:creator>
<foaf:homepage rdf:resource="http://localhost:8080/"/>
<dc:contributor>Mayans, Olga</dc:contributor>
<dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/45025/1/Adams_2-14disd67exh901.pdf"/>
<dc:contributor>Markovic, Marija</dc:contributor>
<dc:creator>Fleming, Jennifer R.</dc:creator>
<dc:contributor>Zhou, Tiankun</dc:contributor>
<dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-02-14T10:01:08Z</dc:date>
<dcterms:issued>2019-04</dcterms:issued>
<dc:contributor>Fleming, Jennifer R.</dc:contributor>
<dc:creator>Zacharchenko, Thomas</dc:creator>
<dcterms:title>Scalable, Non-denaturing Purification of Phosphoproteins Using Ga<sup>3+</sup>-IMAC : N2A and M1M2 Titin Components as Study case</dcterms:title>
<dc:creator>Mayans, Olga</dc:creator>
<dcterms:abstract xml:lang="eng">The purification of phosphorylated proteins in a folded state and in large enough quantity for biochemical or biophysical analysis remains a challenging task. Here, we develop a new implementation of the method of gallium immobilized metal chromatography (Ga<sup>3+</sup>-IMAC) as to permit the selective enrichment of phosphoproteins in the milligram scale and under native conditions using automated FPLC instrumentation. We apply this method to the purification of the UN2A and M1M2 components of the muscle protein titin upon being monophosphorylated in vitro by cAMP-dependent protein kinase (PKA). We found that UN2A is phosphorylated by PKA at its C-terminus in residue S9578 and M1M2 is phosphorylated in its interdomain linker sequence at position T32607. We demonstrate that the Ga<sup>3+</sup>-IMAC method is efficient, economical and suitable for implementation in automated purification pipelines for recombinant proteins. The procedure can be applied both to the selective enrichment and to the removal of phosphoproteins from biochemical samples.</dcterms:abstract>
</rdf:Description>
</rdf:RDF>Interner Vermerk
xmlui.Submission.submit.DescribeStep.inputForms.label.kops_note_fromSubmitter
Prüfungsdatum der Dissertation
Finanzierungsart
Kommentar zur Publikation
Allianzlizenz
Corresponding Authors der Uni Konstanz vorhanden
Internationale Co-Autor:innen
Universitätsbibliographie
Ja
Begutachtet
Ja
