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Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
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Development and characterization of protein kinase B/AKT isoform-specific nanobodies

Tijs Merckaert1,2, Olivier Zwaenepoel1, Kris Gevaert1,2

  • 1Department of Biomolecular Medicine, Faculty of Medicine and Health Sciences, Ghent University, Ghent, Belgium.

Plos One
|October 12, 2020
PubMed

Insights

New nanobodies target specific AKT kinase isoforms (AKT1, AKT2), crucial for cancer. These tools aid research into isoform-specific functions and AKT-PIP3 interactions, advancing cancer therapy development.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Research

Background:

  • Serine/threonine protein kinase AKT is over-activated in cancer, correlating with poor prognosis.
  • AKT is a key regulator in the PI3K/AKT/mTOR pathway, a hallmark of cancer.
  • AKT has three highly homologous but non-redundant isoforms (AKT1, AKT2, AKT3) with context-dependent functions.

Purpose of the Study:

  • To develop novel tools for studying AKT isoform-specific functions.
  • To create isoform-specific inhibitors for AKT1 and AKT2.
  • To investigate the AKT-PIP3 interaction using targeted nanobodies.

Main Methods:

  • Development and characterization of nanobodies targeting AKT1 and AKT2.
  • Assessment of nanobody interaction with specific AKT isoforms.
  • Evaluation of nanobodies' ability to inhibit AKT-PIP3 interaction.

Main Results:

  • Successfully developed and characterized nanobodies specific to AKT1 and AKT2.
  • Obtained nanobodies that interfere with the essential AKT-PIP3 interaction for both isoforms.
  • Demonstrated the potential of nanobodies as tools for isoform-specific AKT research.

Conclusions:

  • Nanobodies offer a non-genetic approach to study AKT1 and AKT2 isoform-specific functions.
  • These nanobodies are valuable tools for understanding AKT signaling in cancer.
  • The developed nanobodies represent a step towards targeted cancer therapies by enabling deeper insights into AKT isoform activity.

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