Crystal structure-guided design of bisubstrate inhibitors and photoluminescent probes for protein kinases of the PIM family

  • We performed an X-ray crystallographic study of complexes of protein kinase PIM-1 with three inhibitors comprising an adenosine mimetic moiety, a linker, and a peptide-mimetic (d-Arg)6 fragment. Guided by the structural models, simplified chemical structures with a reduced number of polar groups and chiral centers were designed. The developed inhibitors retained low-nanomolar potency and possessed remarkable selectivity toward the PIM kinases. The new inhibitors were derivatized with biotin or fluorescent dye Cy5 and then applied for the detection of PIM kinases in biochemical solutions and in complex biological samples. The sandwich assay utilizing a PIM-2-selective detection antibody featured a low limit of quantification (44 pg of active recombinant PIM-2). Fluorescent probes were efficiently taken up by U2OS cells and showed a high extent of co-localization with PIM-1 fused with a fluorescent protein. Overall, the developed inhibitors and derivatives represent versatile chemical tools for studying PIM function in cellular systems in normal and disease physiology.
Author:Olivier E. Nonga, Darja Lavogina, Erki Enkvist, Katrin Kestav, Apirat ChaikuadORCiD, Sarah E. Dixon-Clarke, Alex N. BullockORCiD, Sergei Kopanchuk, Taavi Ivan, Ramesh Ekambaram, Kaido Viht, Stefan KnappORCiD, Asko Uri
Parent Title (English):Molecules
Place of publication:Basel
Document Type:Article
Date of Publication (online):2021/07/19
Date of first Publication:2021/07/19
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2021/11/09
Tag:PIM kinases; adenosine–arginine conjugate; bisubstrate inhibitors; cellular uptake and localization; co-crystal structure; fluorescent probes; protein X-ray crystallography
Issue:14, art. 4353
Page Number:22
First Page:1
Last Page:22
The work was supported by the Estonian Research Council grants PRG454 and PSG230. O.E.N. was supported by the UT ASTRA project Graduate School “Functional materials and technologies” receiving funding from the European Regional Development Fund (OEN). S.E.D.-C. was supported by a NDM Prize Studentship, which was funded by the Medical Research Council and the Nuffield Department of Medicine, University of Oxford. S.Kn. and A.C. are grateful for support by the SGC, a registered charity (no. 1097737) that receives funds from AbbVie, BayerAG, Boehringer Ingelheim, the Canada Foundation for Innovation, Eshelman Institute for Innovation, Genentech, Genome Canada through Ontario Genomics Institute (OGI-196), EU/EFPIA/OICR/McGill/KTH/Diamond, Innovative Medicines Initiative 2 Joint Undertaking (EUbOPEN grant 875510), Janssen, Merck KGaA, Merck & Co, Pfizer, the São Paulo Research Foundation-FAPESP, Takeda and Wellcome (106169/ZZ14/Z). S.Kn. is also grateful for support by the German translational cancer network (DKTK) as well as the Frankfurt Cancer Institute (FCI).
Institutes:Biochemie, Chemie und Pharmazie
Dewey Decimal Classification:6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit
Licence (German):License LogoCreative Commons - Namensnennung 4.0