Related Experiment Video
Updated: Jan 9, 2026
Assaying Protein Kinase Activity with Radiolabeled ATP
Published on: May 26, 2017
The Discovery of Abl Kinase ATPase Activity and Its Implications in the Development of Straightforward Assays
Diego Magno Martins1, Philipe Oliveira Fernandes2, Lucas Almeida Vieira1
1Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais (UFMG), 31270-901 Belo Horizonte, MG, Brazil.
Abstract:
Abelson kinase (Abl) is an enzyme crucial in metabolic pathways, and it is a molecular target for leukemias, especially chronic myeloid leukemia. Despite extensive research on its kinase function and inhibition over the years, there is still considerable room for new discoveries regarding its mechanistic and reactive aspects. We report here, for the first time, the intrinsic ATPase activity of Abl in the absence of peptide substrates. Using quantitative one-dimensional 31P nuclear magnetic resonance spectroscopy, we monitored the conversion of ATP to ADP and inorganic phosphate (Pi) catalyzed by Abl. Furthermore, we demonstrated that the known kinase inhibitors imatinib and dasatinib inhibit ATPase activity. Beyond expanding the biochemical repertoire of Abl, this finding enables a straightforward, substrate-free assay for kinase inhibition, offering new perspectives on kinase catalytic plasticity and laying the groundwork for simplified screening strategies in drug discovery.
Insights
Abelson kinase (Abl) exhibits intrinsic ATPase activity without peptide substrates. This discovery enables a novel, substrate-free assay for kinase inhibitors like imatinib and dasatinib, aiding drug discovery.
Area of Science:
- Biochemistry
- Molecular Biology
- Enzymology
Background:
- Abelson kinase (Abl) is a key enzyme in metabolic pathways.
- Abl is a critical molecular target for leukemias, particularly chronic myeloid leukemia.
- Existing research primarily focuses on Abl's kinase function and inhibition, with limited understanding of its mechanistic and reactive aspects.
Purpose of the Study:
- To investigate the intrinsic ATPase activity of Abelson kinase (Abl) in the absence of peptide substrates.
- To characterize the substrate-free catalytic activity of Abl.
- To establish a novel assay for kinase inhibition screening.
Main Methods:
- Quantitative one-dimensional 31P nuclear magnetic resonance (NMR) spectroscopy was employed.
- The conversion of ATP to ADP and inorganic phosphate (Pi) catalyzed by Abl was monitored.
- The inhibitory effects of imatinib and dasatinib on Abl's ATPase activity were assessed.
Main Results:
- The intrinsic ATPase activity of Abl was demonstrated for the first time, independent of peptide substrates.
- The conversion of ATP to ADP and Pi by Abl was quantitatively measured.
- Established kinase inhibitors imatinib and dasatinib were shown to inhibit Abl's intrinsic ATPase activity.
Conclusions:
- Abelson kinase possesses intrinsic ATPase activity, expanding its known biochemical repertoire.
- This finding facilitates a straightforward, substrate-free assay for evaluating kinase inhibitors.
- The study offers new insights into kinase catalytic plasticity and supports simplified drug discovery screening strategies.
