title: CDC25A-inhibitory RE derivatives bind to pocket adjacent to the catalytic site.
abstract: RE derivatives, which are cell-permeable and non-electrophilic dual-specificity protein phosphatase inhibitors developed in our laboratory, inhibit CDC25A/B non-competitively, as determined by means of kinetic experiments. To identify the binding site of RE derivatives, we designed and synthesized the new probe molecule RE142, having a Michael acceptor functionality for covalent bond formation with the enzyme, a biotin tag to enable enrichment of probe-bound peptide(s), and a chemically cleavable linker to facilitate release of probe-bound peptides from avidin beads. LC-MS analysis indicated that RE142 binds to one of the residues Cys384-Tyr386 of CDC25A, within a pocket adjacent to the catalytic site.
authors: RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
authors: Ayako
authors: A
authors: Tsuchiya
authors: Miwako
authors: M
authors: Asanuma
authors: Go
authors: G
authors: Hirai
authors: Kana
authors: K
authors: Oonuma
authors: Muhammad
authors: M
authors: Muddassar
authors: Eri
authors: E
authors: Nishizawa
authors: Yusuke
authors: Y
authors: Koyama
authors: Yuko
authors: Y
authors: Otani
authors: Kam Y J
authors: KY
authors: Zhang
authors: Mikiko
authors: M
authors: Sodeoka
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry / Q000379:methods; D008956:Models, Chemical; D008958:Models, Molecular; D008968:Molecular Conformation; D015394:Molecular Structure; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity; D020687:cdc25 Phosphatases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D020687:cdc25 Phosphatases