Toward G protein-coupled receptor structure-based drug design using X-ray lasers

Area of Science:

• Structural biology
• Biochemistry
• Pharmacology

Background:

• Structure-based drug design (SBDD) is crucial for optimizing drug leads.
• G protein-coupled receptors (GPCRs) are important drug targets but difficult to crystallize, limiting SBDD.
• Previous crystallization methods for GPCRs were challenging and time-consuming.

Purpose of the Study:

• To develop a rapid method for determining multiple co-crystal structures of GPCRs with various ligands.
• To enable structure-based drug design for GPCRs and other challenging membrane proteins.

Main Methods:

• Utilized serial femtosecond crystallography (SFX) to overcome limitations of large crystal requirements.
• Employed SFX for rapid co-crystal structure determination using submilligram protein quantities.
• Applied the method to the human β2-adrenergic receptor and three additional receptors.

Main Results:

• Successfully determined eight room-temperature co-crystal structures of the human β2-adrenergic receptor with six different ligands.
• Obtained previously unreported co-crystal structures, including those with carvedilol and propranolol.
• Demonstrated the method's generality by successfully testing it on three other GPCR targets.

Conclusions:

• The developed SFX-based method significantly accelerates the determination of GPCR co-crystal structures.
• This approach overcomes major crystallization hurdles, making GPCRs more amenable to SBDD.
• The method holds promise for advancing drug discovery targeting GPCRs and other recalcitrant membrane proteins.