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Segmental Isotopic Labeling of a Central Domain in a Multidomain Protein by Protein Trans-Splicing Using Only One Robust DnaE Intein

Authors

  • Alena E. L. Busche,

    1. Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Frankfurt Macromolecular Complexes (CEF), University of Frankfurt, 60438 Frankfurt (Germany)
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    • These authors contributed equally to this work.

  • A. Sesilja Aranko,

    1. Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki 00014 (Finland), Fax: (+358) 9-1915-9541 http://www.biocenter.helsinki.fi/bi/iwai/
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    • These authors contributed equally to this work.

  • Mehdi Talebzadeh-Farooji Dr.,

    1. Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Frankfurt Macromolecular Complexes (CEF), University of Frankfurt, 60438 Frankfurt (Germany)
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  • Frank Bernhard Dr.,

    1. Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Frankfurt Macromolecular Complexes (CEF), University of Frankfurt, 60438 Frankfurt (Germany)
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  • Volker Dötsch Prof. Dr.,

    1. Institute of Biophysical Chemistry and Center for Biomolecular Magnetic Resonance and Cluster of Excellence Frankfurt Macromolecular Complexes (CEF), University of Frankfurt, 60438 Frankfurt (Germany)
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  • Hideo Iwaï Dr.

    1. Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki, P.O. Box 65, Helsinki 00014 (Finland), Fax: (+358) 9-1915-9541 http://www.biocenter.helsinki.fi/bi/iwai/
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  • We thank the Protein Chemistry Research Group at the Institute of Biotechnology and Dr. Ute Bahr for mass spectrometry analysis. This work was supported by the grants from the Academy of Finland (118385), Sigrid Jusélius Foundation, the Center for Biomolecular Magnetic Resonance (BMRZ), and the Cluster of Excellence Frankfurt (CEF).

Abstract

original image

Der richtige Griff: Mit einem natürlich gespaltenen Intein wurde eine effiziente Dreiwege-Ligationsmethode entwickelt, die ohne Rückfaltungsschritte auskommt. Auf diese Art gelingt die selektive Markierung einer zentralen Domäne in einem Dreidomänenprotein mit NMR-aktiven Isotopen (siehe Bild), was die Untersuchung von Domänen-Domänen-Wechselwirkungen in Mehrdomänenproteinen ermöglicht.

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