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Updated: Jan 19, 2026
Fully Autonomous Characterization and Data Collection from Crystals of Biological Macromolecules
Published on: March 22, 2019
High-resolution crystal structure of human asparagine synthetase enables analysis of inhibitor binding and
Wen Zhu1,2, Ashish Radadiya1, Claudine Bisson3,2
11School of Chemistry, Cardiff University, Cardiff, UK.
Abstract:
Expression of human asparagine synthetase (ASNS) promotes metastatic progression and tumor cell invasiveness in colorectal and breast cancer, presumably by altering cellular levels of L-asparagine. Human ASNS is therefore emerging as a bona fide drug target for cancer therapy. Here we show that a slow-onset, tight binding inhibitor, which exhibits nanomolar affinity for human ASNS in vitro, exhibits excellent selectivity at 10 μM concentration in HCT-116 cell lysates with almost no off-target binding. The high-resolution (1.85 Å) crystal structure of human ASNS has enabled us to identify a cluster of negatively charged side chains in the synthetase domain that plays a key role in inhibitor binding. Comparing this structure with those of evolutionarily related AMP-forming enzymes provides insights into intermolecular interactions that give rise to the observed binding selectivity. Our findings demonstrate the feasibility of developing second generation human ASNS inhibitors as lead compounds for the discovery of drugs against metastasis.
Insights
A novel inhibitor targeting human asparagine synthetase (ASNS) shows high selectivity and nanomolar affinity, offering a promising strategy for developing new anti-metastasis cancer drugs.
Area of Science:
- Biochemistry
- Molecular Biology
- Cancer Research
Background:
- Human asparagine synthetase (ASNS) expression correlates with metastatic progression in colorectal and breast cancers.
- ASNS is a validated drug target for cancer therapy due to its role in L-asparagine metabolism.
Purpose of the Study:
- To characterize a novel, selective inhibitor of human ASNS.
- To elucidate the structural basis for ASNS inhibitor binding and selectivity.
Main Methods:
- In vitro biochemical assays to determine inhibitor affinity and selectivity.
- High-resolution (1.85 Å) crystal structure determination of human ASNS.
- Comparative structural analysis with related AMP-forming enzymes.
Main Results:
- A novel inhibitor demonstrated nanomolar affinity and excellent selectivity for human ASNS.
- The crystal structure revealed key negatively charged residues in the synthetase domain crucial for inhibitor binding.
- Structural comparisons provided insights into the molecular interactions driving binding selectivity.
Conclusions:
- Development of selective human ASNS inhibitors is feasible.
- This study provides a foundation for designing second-generation ASNS inhibitors as anti-metastasis drug leads.
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