Identification of the molecular composition of the 20S proteasome of mouse intestine by high-resolution mass spectrometric proteome analysis

Loading...
Thumbnail Image
Date
2009
Authors
Preywisch, Regina
Youhnovski, Nikolay
Editors
Contact
Journal ISSN
Electronic ISSN
ISBN
Bibliographical data
Publisher
Series
ArXiv-ID
International patent number
Link to the license
EU project number
Project
Open Access publication
Collections
Restricted until
Title in another language
Research Projects
Organizational Units
Journal Issue
Publication type
Journal article
Publication status
Published in
Methods in Molecular Biology ; 564 (2009). - pp. 173-186. - ISSN 1064-3745
Abstract
In the last years, intracellular protein degradation by the proteasome has become a focus area of scientific interest. Here, we describe a proteomics approach for the molecular mapping of the constituents of the proteolytically active core particle, the constitutive 20S proteasome from mouse intestine. In addition to the proteomics workflow widely used for protein isolation, gel electrophoretic separation, in-gel digestion, and UV-MALDI mass spectrometry, high-resolution Fourier transform ion cyclotron resonance mass spectrometry using infrared-MALDI ionisation (IR-MALDI FTICR-MS) has been employed as an efficient method for protein identification by peptide mass fingerprint. The 20S proteasome subunits α1 α7 and β1 β7 were completely and unambiguously identified. In addition to subunits β1 and β2, the corresponding inducible subunits being part of the immuno-proteasome were identified. The subunit β5i was found to completely replace the corresponding constitutive subunit, suggesting a high proteolytic activity of the intestinal proteasome leading to increased production of antigenic peptides. The high mass accuracy in the low ppm range and resolution of FTICR-MS provide direct identifications of individual proteins as mixtures such as components resulting from incomplete electrophoretic separation. In addition, the comparison of UV- and IR-MALDI FTICR-MS may provide details of fragmentation and rearrangement reactions that may occur under UV-MALDI ionisation conditions.
Summary in another language
Subject (DDC)
570 Biosciences, Biology
Keywords
20S-Proteasome,High-resolution FTICR mass spectrometry,Proteome analysis,Proteasome constituents,UV- and IR-MALDI ionisation
Conference
Review
undefined / . - undefined, undefined. - (undefined; undefined)
Cite This
ISO 690WEBER, Reinhold, Regina PREYWISCH, Nikolay YOUHNOVSKI, Marcus GRÖTTRUP, Michael PRZYBYLSKI, 2009. Identification of the molecular composition of the 20S proteasome of mouse intestine by high-resolution mass spectrometric proteome analysis. In: Methods in Molecular Biology. 564, pp. 173-186. ISSN 1064-3745. Available under: doi: 10.1007/978-1-60761-157-8_10
BibTex
@article{Weber2009Ident-10054,
  year={2009},
  doi={10.1007/978-1-60761-157-8_10},
  title={Identification of the molecular composition of the 20S proteasome of mouse intestine by high-resolution mass spectrometric proteome analysis},
  volume={564},
  issn={1064-3745},
  journal={Methods in Molecular Biology},
  pages={173--186},
  author={Weber, Reinhold and Preywisch, Regina and Youhnovski, Nikolay and Gröttrup, Marcus and Przybylski, 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/123456789/10054">
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dc:creator>Przybylski, Michael</dc:creator>
    <dcterms:abstract xml:lang="eng">In the last years, intracellular protein degradation by the proteasome has become a focus area of scientific interest. Here, we describe a proteomics approach for the molecular mapping of the constituents of the proteolytically active core particle, the constitutive 20S proteasome from mouse intestine. In addition to the proteomics workflow widely used for protein isolation, gel electrophoretic separation, in-gel digestion, and UV-MALDI mass spectrometry, high-resolution Fourier transform ion cyclotron resonance mass spectrometry using infrared-MALDI ionisation (IR-MALDI FTICR-MS) has been employed as an efficient method for protein identification by peptide mass fingerprint. The 20S proteasome subunits α1 α7 and β1 β7 were completely and unambiguously identified. In addition to subunits β1 and β2, the corresponding inducible subunits being part of the immuno-proteasome were identified. The subunit β5i was found to completely replace the corresponding constitutive subunit, suggesting a high proteolytic activity of the intestinal proteasome leading to increased production of antigenic peptides. The high mass accuracy in the low ppm range and resolution of FTICR-MS provide direct identifications of individual proteins as mixtures such as components resulting from incomplete electrophoretic separation. In addition, the comparison of UV- and IR-MALDI FTICR-MS may provide details of fragmentation and rearrangement reactions that may occur under UV-MALDI ionisation conditions.</dcterms:abstract>
    <dc:contributor>Youhnovski, Nikolay</dc:contributor>
    <dc:rights>terms-of-use</dc:rights>
    <dc:contributor>Weber, Reinhold</dc:contributor>
    <dc:creator>Youhnovski, Nikolay</dc:creator>
    <dc:language>eng</dc:language>
    <dc:creator>Preywisch, Regina</dc:creator>
    <dc:format>application/pdf</dc:format>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:creator>Weber, Reinhold</dc:creator>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/10054/1/Przybylski_Michael2009_h.pdf"/>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/10054/1/Przybylski_Michael2009_h.pdf"/>
    <dc:contributor>Przybylski, Michael</dc:contributor>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-24T18:16:16Z</dc:date>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/>
    <dc:contributor>Preywisch, Regina</dc:contributor>
    <dcterms:bibliographicCitation>First publ. in: Methods in Molecular Biology ; 564 (2009). - S. 173-186</dcterms:bibliographicCitation>
    <dcterms:title>Identification of the molecular composition of the 20S proteasome of mouse intestine by high-resolution mass spectrometric proteome analysis</dcterms:title>
    <dc:creator>Gröttrup, Marcus</dc:creator>
    <dcterms:issued>2009</dcterms:issued>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/10054"/>
    <dc:contributor>Gröttrup, Marcus</dc:contributor>
  </rdf:Description>
</rdf:RDF>
Internal note
xmlui.Submission.submit.DescribeStep.inputForms.label.kops_note_fromSubmitter
Contact
URL of original publication
Test date of URL
Examination date of dissertation
Method of financing
Comment on publication
Alliance license
Corresponding Authors der Uni Konstanz vorhanden
International Co-Authors
Bibliography of Konstanz
Yes
Refereed