Stabilization of Intrinsically Disordered DKK2 Protein by Fusion to RNA-Binding Domain

International Journal of Molecular Sciences

ByungKyu Lee¹  BaikL. Seong¹  Young-Guen Kwon²  Jonghyo Lim²  SoonBin Kwon³  Ahyun Son³  HyeMin Lee³  Kyun-Hwan Kim⁴  DooHyun Kim⁴  JinSun Kang⁵  YoungHo Byun⁵ 

[1] D Institute, ProCell Therapeutics, Inc., Ace-Twin Tower II, Guro3-dong, Guro-gu, Seoul 08381, Korea;Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 03722, Korea;Department of Biotechnology, College of Life Sciences and Biotechnology, Yonsei University, Seoul 03722, Korea;Department of Pharmacology, and Center for Cancer Research and Diagnostic Medicine, IBST, School of Medicine, Konkuk University, Seoul 05030, Korea;;ProCell R&

关键词: DKK2;    IDPs;    RBPs;    Wnt signaling;    angiogenesis;   

DOI  :  10.3390/ijms20112847

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown