Selective inhibition of the kinase DYRK1A by targeting its folding process

Isao Kii; Yuto Sumida; Toshiyasu Goto; Rie Sonamoto; Yukiko Okuno; Suguru Yoshida; Tomoe Kato-Sumida; Yuka Koike; Minako Abe; Yosuke Nonaka; Teikichi Ikura; Nobutoshi Ito; Hiroshi Shibuya; Takamitsu Hosoya; Masatoshi Hagiwara

doi:10.1038/ncomms11391

Selective inhibition of the kinase DYRK1A by targeting its folding process

Nat Commun. 2016 Apr 22:7:11391. doi: 10.1038/ncomms11391.

Authors

Isao Kii^{1

2}, Yuto Sumida³, Toshiyasu Goto⁴, Rie Sonamoto¹, Yukiko Okuno¹, Suguru Yoshida³, Tomoe Kato-Sumida³, Yuka Koike^{1

2}, Minako Abe⁵, Yosuke Nonaka⁵, Teikichi Ikura⁵, Nobutoshi Ito⁵, Hiroshi Shibuya⁴, Takamitsu Hosoya³, Masatoshi Hagiwara¹

Affiliations

¹ Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.
² Pathophysiological and Health Science Team, Imaging Application Group, Division of Bio-Function Dynamics Imaging, RIKEN Center for Life Science Technologies, 6-7-3 Minatojima-minamimachi, Chuo-ku, Kobe 650-0047, Japan.
³ Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Tokyo 101-0062, Japan.
⁴ Department of Molecular Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Tokyo 113-8510, Japan.
⁵ Department of Structural Biology, Graduate School of Medical and Dental Sciences, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Tokyo 113-8510, Japan.

Abstract

Autophosphorylation of amino-acid residues is part of the folding process of various protein kinases. Conventional chemical screening of mature kinases has missed inhibitors that selectively interfere with the folding process. Here we report a cell-based assay that evaluates inhibition of a kinase at a transitional state during the folding process and identify a folding intermediate-selective inhibitor of dual-specificity tyrosine-phosphorylation-regulated kinase 1A (DYRK1A), which we refer to as FINDY. FINDY suppresses intramolecular autophosphorylation of Ser97 in DYRK1A in cultured cells, leading to its degradation, but does not inhibit substrate phosphorylation catalysed by the mature kinase. FINDY also suppresses Ser97 autophosphorylation of recombinant DYRK1A, suggesting direct inhibition, and shows high selectivity for DYRK1A over other DYRK family members. In addition, FINDY rescues DYRK1A-induced developmental malformations in Xenopus laevis embryos. Our study demonstrates that transitional folding intermediates of protein kinases can be targeted by small molecules, and paves the way for developing novel types of kinase inhibitors.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Biological Assay*
Biomarkers / metabolism
Cantharidin / pharmacology
Dyrk Kinases
Embryo, Nonmammalian
Gene Expression Regulation, Developmental
HEK293 Cells
Humans
Marine Toxins
Molecular Sequence Data
Okadaic Acid / pharmacology
Oxazoles / pharmacology
Phosphorylation / drug effects
Plasmids / chemistry
Plasmids / metabolism
Protein Folding / drug effects*
Protein Kinase Inhibitors / chemistry
Protein Kinase Inhibitors / pharmacology*
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Protein Serine-Threonine Kinases / chemistry
Protein Serine-Threonine Kinases / genetics
Protein Serine-Threonine Kinases / metabolism
Protein-Tyrosine Kinases / antagonists & inhibitors*
Protein-Tyrosine Kinases / chemistry
Protein-Tyrosine Kinases / genetics
Protein-Tyrosine Kinases / metabolism
Recombinant Proteins
Sequence Alignment
Thiazoles / chemistry
Thiazoles / pharmacology*
Transfection
Xenopus laevis / embryology

Substances

Biomarkers
FINDY compound
Marine Toxins
Oxazoles
Protein Kinase Inhibitors
Recombinant Proteins
Thiazoles
Okadaic Acid
calyculin A
Protein-Tyrosine Kinases
Protein Serine-Threonine Kinases
Cantharidin