Stabilization of intrinsically disordered DKK2 protein by fusion to RNA-binding domain

Hye Min Lee, Soon Bin Kwon, Ahyun Son, Doo Hyun Kim, Kyun Hwan Kim, Jonghyo Lim, Young Guen Kwon, Jin Sun Kang, Byung Kyu Lee, Young Ho Byun, Baik L. Seong

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Abstract

Intrinsic disorders are a common feature of hub proteins in eukaryotic interactomes controlling the signaling pathways. The intrinsically disordered proteins (IDPs) are prone to misfolding, and maintaining their functional stability remains a major challenge in validating their therapeutic potentials. Considering that IDPs are highly enriched in RNA-binding proteins (RBPs), here we reasoned and confirmed that IDPs could be stabilized by fusion to RBPs. Dickkopf2 (DKK2), Wnt antagonist and a prototype IDP, was fused with lysyl-tRNA synthetase (LysRS), with or without the fragment crystallizable (Fc) domain of an immunoglobulin and expressed predominantly as a soluble form from a bacterial host. The functional competence was confirmed by in vitro Wnt signaling reporter and tube formation in human umbilical vein endothelial cells (HUVECs) and in vivo Matrigel plug assay. The removal of LysRS by site-specific protease cleavage prompted the insoluble aggregation, confirming that the linkage to RBP chaperones the functional competence of IDPs. While addressing to DKK2 as a key modulator for cancer and ischemic vascular diseases, our results suggest the use of RBPs as stabilizers of disordered proteinaceous materials for acquiring and maintaining the structural stability and functional competence, which would impact the druggability of a variety of IDPs from human proteome.

Original languageEnglish
Article number2847
JournalInternational journal of molecular sciences
Volume20
Issue number11
DOIs
Publication statusPublished - 2019 Jun 1

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All Science Journal Classification (ASJC) codes

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Lee, H. M., Kwon, S. B., Son, A., Kim, D. H., Kim, K. H., Lim, J., Kwon, Y. G., Kang, J. S., Lee, B. K., Byun, Y. H., & Seong, B. L. (2019). Stabilization of intrinsically disordered DKK2 protein by fusion to RNA-binding domain. International journal of molecular sciences, 20(11), [2847]. https://doi.org/10.3390/ijms20112847