Improving coiled coil stability while maintaining specificity by a bacterial hitchhiker selection system

Kuekenshoener 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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Abstract
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.
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Kuekenshoener 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.
Kuekenshoener, 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.
Kuekenshoener, 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.
Kuekenshoener, 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. Kuekenshoener, 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.
Kuekenshoener, 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).
Kuekenshoener, 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.
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Controlling leucine-zipper partner recognition in cells through modification of a-g interactions.
Azuma Y, Kukenshoner T, Ma G, Yasunaga J, Imanishi M, Tanaka G, Nakase I, Maruno T, Kobayashi Y, Arndt KM, Matsuoka M, Futaki S., Chem. Commun. (Camb.) 50(48), 2014
PMID: 24803110

63 References

Data provided by Europe PubMed Central.

The semidominant Mi(b) mutation identifies a role for the HLH domain in DNA binding in addition to its role in protein dimerization.
Steingrimsson E, Nii A, Fisher DE, Ferre-D'Amare AR, McCormick RJ, Russell LB, Burley SK, Ward JM, Jenkins NA, Copeland NG., EMBO J. 15(22), 1996
PMID: 8947051
Melanocytes and the microphthalmia transcription factor network.
Steingrimsson E, Copeland NG, Jenkins NA., Annu. Rev. Genet. 38(), 2004
PMID: 15568981
Waardenburg syndrome type 2 caused by mutations in the human microphthalmia (MITF) gene.
Tassabehji M, Newton VE, Read AP., Nat. Genet. 8(3), 1994
PMID: 7874167
TFE3, a transcription factor homologous to microphthalmia, is a potential transcriptional activator of tyrosinase and TyrpI genes.
Verastegui C, Bertolotto C, Bille K, Abbe P, Ortonne JP, Ballotti R., Mol. Endocrinol. 14(3), 2000
PMID: 10707962
Automated structure solution with autoSHARP.
Vonrhein C, Blanc E, Roversi P, Bricogne G., Methods Mol. Biol. 364(), 2007
PMID: 17172768
Extended knobs-into-holes packing in classical and complex coiled-coil assemblies.
Walshaw J, Woolfson DN., J. Struct. Biol. 144(3), 2003
PMID: 14643203
Potent D-peptide inhibitors of HIV-1 entry.
Welch BD, VanDemark AP, Heroux A, Hill CP, Kay MS., Proc. Natl. Acad. Sci. U.S.A. 104(43), 2007
PMID: 17942675
Reengineering natural design by rational design and in vivo library selection: the HLH subdomain in bHLHZ proteins is a unique requirement for DNA-binding function.
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

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