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  An automated in vitro protein folding screen applied to a human dynactin subunit

Scheich, C., Niesen, F. H., Seckler, R., & Büssow, K. (2004). An automated in vitro protein folding screen applied to a human dynactin subunit. Protein Science, 13(2), 370-380. doi:10.1110/ps.03304604.

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Genre: Zeitschriftenartikel
Alternativer Titel : Prot Sci

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 Urheber:
Scheich, Christoph1, Autor
Niesen, Frank H., Autor
Seckler, Robert, Autor
Büssow, Konrad2, Autor           
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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Schlagwörter: Folding screen; refolding; inclusion bodies; hydrophobic interaction chromatography; tryptophan fluorescence spectroscopy; human dynactin; structural genomics
 Zusammenfassung: The preparation of proteins for structural and functional analysis using the Escherichia coli expression system is often hampered by the formation of insoluble intracellular protein aggregates (inclusion bodies). Transferring those proteins into their native states by in vitro protein folding requires screening for the best buffer conditions and suitable additives. However, it is difficult to assess the success of such a screen if no biological assay is available. We established a fully automated folding screen and a system to detect folded protein that is based on analytical hydrophobic interaction chromatography and tryptophan fluorescence spectroscopy. The system was evaluated with two model enzymes (carbonic anhydrase II and malate dehydrogenase), and was successfully applied to the folding of the p22 subunit of human dynactin, which is expressed in inclusion bodies in E. coli. The described screen allows for high-throughput folding analysis of inclusion body proteins for structural and functional analyses.

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Sprache(n): eng - English
 Datum: 2004-02
 Publikationsstatus: Erschienen
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Titel: Protein Science
  Alternativer Titel : Prot Sci
Genre der Quelle: Zeitschrift
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Ort, Verlag, Ausgabe: -
Seiten: - Band / Heft: 13 (2) Artikelnummer: - Start- / Endseite: 370 - 380 Identifikator: ISSN: 0961-8368