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Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Pim kinase substrate identification and specificity

Charline Peng1, Axel Knebel, Nick A Morrice

  • 1Boehringer Ingelheim Pharmaceuticals, Inc. 900 Ridgebury Road, Ridgefield, CT 06877, USA.

Journal of Biochemistry
|January 20, 2007
PubMed

Insights

Researchers identified a new Pim kinase recognition sequence (RXRHXS) crucial for lymphomagenesis and cell survival. This discovery aids in developing sensitive assays for screening Pim kinase modulators.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Oncology

Background:

  • The Pim kinase family plays a role in lymphomagenesis and cell survival.
  • Known substrates for Pim kinases are limited and not well-understood.

Purpose of the Study:

  • To identify novel substrates of Pim-2 kinase using the KESTREL approach.
  • To characterize the consensus phosphorylation motif recognized by Pim kinases.

Main Methods:

  • Utilized the Kinase Substrate Tracking and Elucidation (KESTREL) method.
  • Performed sequence analysis of identified substrates (eIF4B, API-5) and known substrates (BAD).
  • Synthesized peptides based on the derived consensus sequence for phosphorylation assays.

Main Results:

  • Identified eukaryotic initiation factor 4B (eIF4B) and apoptosis inhibitor 5 (API-5) as novel Pim-2 substrates.
  • Determined a novel Pim kinase consensus phosphorylation sequence: RXRHXS.
  • Demonstrated that Pim-1 and Pim-2 phosphorylate the RXRHXS sequence 20-fold more efficiently than previously known sites.

Conclusions:

  • The discovery of the RXRHXS consensus sequence enhances understanding of Pim kinase substrate specificity.
  • This finding facilitates the development of improved peptide-based assays for screening Pim kinase inhibitors.
  • The identified substrates and consensus sequence offer new insights into Pim kinase-mediated signaling in cancer and cell survival.

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