Surface-mutagenesis strategies to enable structural biology crystallization platforms.

Area of Science:

• Structural biology
• Biochemistry
• Drug discovery

Background:

• Protein crystallography is crucial for drug discovery, requiring rapid structure determination.
• Identifying suitable protein variants and surface mutations is key for successful crystallization.
• Existing methods may not always yield crystals suitable for structural analysis.

Purpose of the Study:

• To present case studies of rationally designed surface modifications for protein crystallization.
• To demonstrate the effectiveness of these modifications in obtaining well-diffracting crystals.
• To analyze crystal packing to understand the role of mutations in promoting crystallization.

Main Methods:

• Rational design of surface mutations on target proteins.
• Establishment of crystallization conditions for protein kinases (Aurora-C, IRAK4, BUB1) and KRAS-SOS1 complex.
• Analysis of crystal packing to elucidate the impact of mutations.

Main Results:

• Successfully obtained well-diffracting crystals for Aurora-C, IRAK4, BUB1, and KRAS-SOS1 complex through surface modifications.
• Identified specific surface mutations that promoted crystallization.
• Crystal packing analysis provided insights into how mutations facilitated crystal formation.

Conclusions:

• Rationally designed surface modifications are effective in establishing robust crystallization systems for drug discovery targets.
• This strategy accelerates the structure-guided inhibitor optimization process.
• The presented approaches are routinely used to support hit-to-lead and lead-optimization projects.