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Development of a high-throughput crystal structure-determination platform for JAK1 using a novel metal-chelator
Nicole L Caspers1, Seungil Han1, Francis Rajamohan1
1Structural Biology, Pfizer Inc., Eastern Point Road, Groton, CT 06340, USA.
Acta Crystallographica. Section F, Structural Biology Communications
|November 10, 2016
Summary
Researchers crystallized the JAK1 kinase domain using a novel method. This technique facilitates ligand exchange, aiding structure-based drug design for protein kinases.
Area of Science:
- Biochemistry
- Structural Biology
- Drug Discovery
Background:
- Crystallization of protein kinases is crucial for structure-based drug design.
- The ATP-binding site often requires a stabilizing ligand for successful crystallization.
- Janus kinase 1 (JAK1) is a key target in various diseases.
Purpose of the Study:
- To develop a method for facile ligand exchange in protein kinase crystals.
- To enable structure-based drug design by allowing the introduction of diverse ligands.
- To overcome challenges in crystallizing kinases with tightly bound native ligands.
Main Methods:
- Co-crystallization of the phosphorylated JAK1 kinase domain with Mg2+-ADP.
- Utilizing EDTA to chelate magnesium and promote ADP release.
- Exchange of ADP with ATP-competitive small-molecule ligands.
Main Results:
- Successfully generated crystals of the JAK1 kinase domain bound to Mg2+-ADP.
- Demonstrated that EDTA treatment effectively dislodges the Mg2+-ADP complex.
- Showed facile exchange of ADP with various ATP-competitive ligands in the crystal structure.
Conclusions:
- A novel crystallization and ligand-exchange method was established for the JAK1 kinase domain.
- This technique overcomes the recalcitrance of native ligands in the ATP-binding site.
- The procedure holds potential for structure-based drug design of other protein kinases.
