Selective inhibition of the kinase DYRK1A by targeting its folding process.
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.
Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.|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.
Isao
I
Kii
Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Tokyo 101-0062, Japan.
Yuto
Y
Sumida
Department of Molecular Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Tokyo 113-8510, Japan.
Toshiyasu
T
Goto
Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.
Rie
R
Sonamoto
Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.
Yukiko
Y
Okuno
Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Tokyo 101-0062, Japan.
Suguru
S
Yoshida
Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Tokyo 101-0062, Japan.
Tomoe
T
Kato-Sumida
Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.|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.
Yuka
Y
Koike
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.
Minako
M
Abe
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.
Yosuke
Y
Nonaka
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.
Teikichi
T
Ikura
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.
Nobutoshi
N
Ito
Department of Molecular Cell Biology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45, Yushima, Tokyo 113-8510, Japan.
Hiroshi
H
Shibuya
Laboratory of Chemical Bioscience, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, 2-3-10 Kanda-Surugadai, Tokyo 101-0062, Japan.
Takamitsu
T
Hosoya
Department of Anatomy and Developmental Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Kyoto 606-8501, Japan.
Masatoshi
M
Hagiwara
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D015415:Biomarkers / Q000378:metabolism; D002193:Cantharidin / Q000494:pharmacology; D004625:Embryo, Nonmammalian; D018507:Gene Expression Regulation, Developmental; D057809:HEK293 Cells; D006801:Humans; D008387:Marine Toxins; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D010080:Oxazoles / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D017510:Protein Folding / Q000187:drug effects*; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins; D016415:Sequence Alignment; D013844:Thiazoles / Q000737:chemistry / Q000494:pharmacology*; D014162:Transfection; D014982:Xenopus laevis / Q000196:embryology; D000097603:Dyrk Kinases
D015415:Biomarkers; D002193:Cantharidin; D008387:Marine Toxins; D019319:Okadaic Acid; D010080:Oxazoles; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases; D011994:Recombinant Proteins; D013844:Thiazoles; D000097603:Dyrk Kinases; C000610374:FINDY compound; C059041:calyculin A