title: Bisubstrate inhibitor approach for targeting mitotic kinase Haspin.
abstract: During the past decade, the basophilic atypical kinase Haspin has emerged as a key player in mitosis responsible for phosphorylation of Thr3 residue of histone H3. Here, we report the construction of conjugates comprising an aromatic fragment targeted to the ATP-site of Haspin and a peptide mimicking the N-terminus of histone H3. The combination of effective solid phase synthesis procedures and a high throughput binding/displacement assay with fluorescence anisotropy readout afforded the development of inhibitors with remarkable subnanomolar affinity toward Haspin. The selectivity profiles of novel conjugates were established by affinity studies with a model basophilic kinase (catalytic subunit of cAMP-dependent protein kinase) and by a commercial 1-point inhibition assay with 43 protein kinases.
authors.affiliation: University of Tartu , Institute of Chemistry, Ravila 14A, Tartu 50411, Estonia.
authors.forename: Katrin
authors.initials: K
authors.lastname: Kestav
authors.forename: Darja
authors.initials: D
authors.lastname: Lavogina
authors.forename: Gerda
authors.initials: G
authors.lastname: Raidaru
authors.forename: Apirat
authors.initials: A
authors.lastname: Chaikuad
authors.forename: Stefan
authors.initials: S
authors.lastname: Knapp
authors.forename: Asko
authors.initials: A
authors.lastname: Uri
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D006657:Histones / Q000737:chemistry; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D008938:Mitosis; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry* / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D010455:Peptides; D047428:Protein Kinase Inhibitors; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; C530656:HASPIN protein, human; D017346:Protein Serine-Threonine Kinases