Slowly on, Slowly off: Bisubstrate-Analogue Conjugates of 5-Iodotubercidin and Histone H3 Peptide Targeting Protein Kinase Haspin.
The atypical protein kinase haspin is a key player in mitosis by catalysing the phosphorylation of Thr3 in histone H3, and thus ensuring the normal function of the chromosomal passenger complex. Here, we report the development of bisubstrate-analogue inhibitors targeting haspin. The compounds were constructed by linking 5-iodotubercidin to the N terminus of histone H3 peptide. The new conjugates show high affinity (sub-nanomolar KD) towards haspin as well as slow kinetics of association and dissociation (residence time of several hours). This reflects a unique binding mode and translated into improved selectivity. The latter was confirmed in a biochemical binding/displacement assay with a panel of ten protein kinases, in a thermal shift assay with off-targets of 5-iodotubercidin (adenosine kinase and the Cdc2-like kinase family) and in assay with spiked HeLa cell lysate.
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Katrin
K
Kestav
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Kaido
K
Viht
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Anton
A
Konovalov
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Erki
E
Enkvist
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Asko
A
Uri
Institute of Chemistry, University of Tartu, Ravila 14A, 50411, Tartu, Estonia.
Darja
D
Lavogina
D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006657:Histones / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry* / Q000494:pharmacology; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D013696:Temperature; D014372:Tubercidin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology
C114557:5-carboxytetramethylrhodamine succinimidyl ester; D005456:Fluorescent Dyes; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; D012235:Rhodamines; C034214:5-iodotubercidin; C530656:HASPIN protein, human; D017346:Protein Serine-Threonine Kinases; D014372:Tubercidin
5-iodotubercidin; association/dissociation kinetics; haspin; histone H3; inhibitors; iodine