Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system
Kükenshöner T, Wohlwend D, Niemoeller C, Dondapati P, Speck J, Adeniran AV, Nieth A, Gerhardt S, Einsle O, Müller K, Arndt KM (2014)
Journal of Structural Biology 186(3): 335-348.
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Autor*in
Kükenshöner, Tim;
Wohlwend, Daniel;
Niemoeller, Christoph;
Dondapati, Padmarupa;
Speck, Janina;
Adeniran, Adebola V.;
Nieth, Anita;
Gerhardt, Stefan;
Einsle, Oliver;
Müller, KristianUniBi ;
Arndt, Katja M.
Abstract / Bemerkung
The design and selection of peptides targeting cellular proteins is challenging and often yields candidates with undesired properties. Therefore we deployed a new selection system based on the twin-arginine translocase (TAT) pathway of Escherichia coli, named hitchhiker translocation (HiT) selection. A pool of alpha-helix encoding sequences was designed and selected for interference with the coiled coil domain (CC) of a melanoma-associated basic-helix-loop-helix-leucine-zipper (bHLHLZ) protein, the microphthalmia associated transcription factor (MITF). One predominant sequence (iM10) was enriched during selection and showed remarkable protease resistance, high solubility and thermal stability while maintaining its specificity. Furthermore, it exhibited nanomolar range affinity towards the target peptide. A mutation screen indicated that target-binding helices of increased homodimer stability and improved expression rates were preferred in the selection process. The crystal structure of the iM10/MITF-CC heterodimer (2.1 angstrom) provided important structural insights and validated our design predictions. Importantly, iM10 did not only bind to the MITF coiled coil, but also to the markedly more stable HLHLZ domain of MITF. Characterizing the selected variants of the semi-rational library demonstrated the potential of the innovative bacterial selection approach. (C) 2014 Elsevier Inc. All rights reserved.
Stichworte
Microphthalmia;
associated transcription factor;
Basic helix-loop-helix leucine zipper;
translocation pathway;
Twin arginine;
Selection and design;
Coiled coils
Erscheinungsjahr
2014
Zeitschriftentitel
Journal of Structural Biology
Band
186
Ausgabe
3
Seite(n)
335-348
ISSN
1047-8477
Page URI
https://pub.uni-bielefeld.de/record/2684271
Zitieren
Kükenshöner T, Wohlwend D, Niemoeller C, et al. Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system. Journal of Structural Biology. 2014;186(3):335-348.
Kükenshöner, T., Wohlwend, D., Niemoeller, C., Dondapati, P., Speck, J., Adeniran, A. V., Nieth, A., et al. (2014). Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system. Journal of Structural Biology, 186(3), 335-348. doi:10.1016/j.jsb.2014.03.002
Kükenshöner, Tim, Wohlwend, Daniel, Niemoeller, Christoph, Dondapati, Padmarupa, Speck, Janina, Adeniran, Adebola V., Nieth, Anita, et al. 2014. “Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system”. Journal of Structural Biology 186 (3): 335-348.
Kükenshöner, T., Wohlwend, D., Niemoeller, C., Dondapati, P., Speck, J., Adeniran, A. V., Nieth, A., Gerhardt, S., Einsle, O., Müller, K., et al. (2014). Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system. Journal of Structural Biology 186, 335-348.
Kükenshöner, T., et al., 2014. Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system. Journal of Structural Biology, 186(3), p 335-348.
T. Kükenshöner, et al., “Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system”, Journal of Structural Biology, vol. 186, 2014, pp. 335-348.
Kükenshöner, T., Wohlwend, D., Niemoeller, C., Dondapati, P., Speck, J., Adeniran, A.V., Nieth, A., Gerhardt, S., Einsle, O., Müller, K., Arndt, K.M.: Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system. Journal of Structural Biology. 186, 335-348 (2014).
Kükenshöner, Tim, Wohlwend, Daniel, Niemoeller, Christoph, Dondapati, Padmarupa, Speck, Janina, Adeniran, Adebola V., Nieth, Anita, Gerhardt, Stefan, Einsle, Oliver, Müller, Kristian, and Arndt, Katja M. “Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system”. Journal of Structural Biology 186.3 (2014): 335-348.
Daten bereitgestellt von European Bioinformatics Institute (EBI)
PDB
1 Eintrag gefunden, die diesen Artikel zitieren
x-ray diffraction (PDB: 4c7n)
Protein structure name: crystal structure of the synthetic peptide im10 in complex with the coiled-coil region of mitf
Public wwPDB file in PDB format
Protein structure name: crystal structure of the synthetic peptide im10 in complex with the coiled-coil region of mitf
Public wwPDB file in PDB format
UNIPROT
1 Eintrag gefunden, die diesen Artikel zitieren
Microphthalmia-associated transcription factor (UNIPROT: O75030)
Organism: Homo sapiens
Download in FASTA format
Organism: Homo sapiens
Download in FASTA format
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Xu J, De Jong AT, Chen G, Chow HK, Damaso CO, Schwartz Mittelman A, Shin JA., Protein Eng. Des. Sel. 23(5), 2010
PMID: 20086039
Xu J, De Jong AT, Chen G, Chow HK, Damaso CO, Schwartz Mittelman A, Shin JA., Protein Eng. Des. Sel. 23(5), 2010
PMID: 20086039
Photocontrol of coiled-coil proteins in living cells.
Zhang F, Timm KA, Arndt KM, Woolley GA., Angew. Chem. Int. Ed. Engl. 49(23), 2010
PMID: 20419720
Zhang F, Timm KA, Arndt KM, Woolley GA., Angew. Chem. Int. Ed. Engl. 49(23), 2010
PMID: 20419720
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