Crystal Structure-Guided Design of Bisubstrate Inhibitors and Photoluminescent Probes for Protein Kinases of the PIM Family

Overview

Journal Molecules

Publisher MDPI

Specialty Biology

Date 2021 Jul 24

PMID 34299628

Citations 2

Authors

Olivier E Nonga

Darja Lavogina

Erki Enkvist

Katrin Kestav

Apirat Chaikuad

Sarah E Dixon-Clarke

Alex N Bullock

Sergei Kopanchuk

Taavi Ivan

Ramesh Ekambaram

Kaido Viht

Stefan Knapp

Asko Uri

Affiliations

Soon will be listed here.

Abstract

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) 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.

Citing Articles

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Picomolar fluorescent probes for compound affinity determination to carbonic anhydrase IX expressed in live cancer cells.

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