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Updated: Sep 24, 2025
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A Semi-High-Throughput Adaptation of the NADH-Coupled ATPase Assay for Screening Small Molecule Inhibitors
Published on: August 17, 2019
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Methods to assess small molecule allosteric modulators of the STRAD pseudokinase
Tingting Qing1, Jin Liu1, Fen Liu1
1Chempartner Co, Ltd., Shanghai, China; Chempartner Co, Ltd., South San Francisco, CA, United States.
Methods in Enzymology
|May 7, 2022
Summary
Developing small molecule modulators for pseudokinases (PSKs) is crucial. This study guides STRAD pseudokinase screening methods to modulate LKB1 kinase activity, favoring direct activity assays for reliable allosteric modulator development.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Pseudokinases (PSKs) play significant biological roles.
- Allosteric modulation of PSKs is a key therapeutic strategy.
- STRAD pseudokinase regulates LKB1 tumor suppressor kinase activity.
Purpose of the Study:
- To provide methods for medium throughput screening of STRAD pseudokinase ligands.
- To assess the impact of ligands on LKB1 kinase activity.
- To guide the development of trans-acting allosteric modulators.
Main Methods:
- Medium throughput screening assays.
- Targeting the ATP binding pocket of STRAD pseudokinase.
- Direct measurement of LKB1 kinase activity.
- Indirect assessment methods for modulator activity.
Main Results:
- Identification of STRAD ligands.
- Evaluation of ligand impact on LKB1 kinase activity.
- Demonstrated limitations of indirect assessment methods.
Conclusions:
- Direct measurement of LKB1 kinase activity is preferred for screening STRAD ligands.
- Indirect methods have limitations in developing trans-acting allosteric modulators.
- This work provides guidance for pseudokinase-targeted drug discovery.
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