Room-temperature X-ray data collection enabled the structural determination of statin-bound CYP105A1

Area of Science:

• Biochemistry
• Structural Biology
• Enzymology

Background:

• Streptomyces griseolus CYP105A1 is a versatile monooxygenase with broad substrate specificity.
• Previous work showed CYP105A1 and its mutants metabolize Vitamin D3, NSAIDs, and statins.
• The R84A mutant demonstrates significant activity against Vitamin D3, NSAIDs, and statins.

Purpose of the Study:

• To elucidate the structural basis of statin binding to CYP105A1.
• To overcome challenges in obtaining high-resolution complex structures of CYP105A1 with statins.

Main Methods:

• Acquisition of complex structures using room-temperature data collection with a conventional capillary method for mevastatin and simvastatin.
• Analysis of cryo-crystallography data and comparison with room-temperature data.

Main Results:

• Successfully obtained complex structures of CYP105A1 with mevastatin and simvastatin at room temperature.
• Observed that cryo-crystals exhibit reduced unit-cell dimensions and increased symmetry interactions.
• Identified cis-trans conversion of the Pro142-Thr143 peptide bond and conformational changes in statin-binding residues within cryo-crystals.

Conclusions:

• Increased symmetry interactions in cryo-crystals likely cause statin dissociation from the active site.
• Room-temperature crystallography is a viable method for studying CYP105A1-statin complexes.
• Structural insights into statin binding are crucial for understanding enzyme-drug interactions.