Toward G protein-coupled receptor structure-based drug design using X-ray lasers
Andrii Ishchenko,
Benjamin Stauch,
Gye Won Han,
Alexander Batyuk,
Anna Shiriaeva,
Chufeng Li,
Nadia Zatsepin,
Uwe Weierstall,
Wei Liu,
Eriko Nango,
Takanori Nakane,
Rie Tanaka,
Kensuke Tono,
Yasumasa Joti,
So Iwata,
Isabel Moraes,
Cornelius Gati,
Vadim Cherezov
Affiliations
Andrii Ishchenko
Bridge Institute, Departments of Chemistry and Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA
Benjamin Stauch
Bridge Institute, Departments of Chemistry and Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA
Gye Won Han
Bridge Institute, Departments of Chemistry and Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA
Alexander Batyuk
Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025, USA
Anna Shiriaeva
Bridge Institute, Departments of Chemistry and Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA
Chufeng Li
Department of Physics, Arizona State University, Tempe, AZ 85287, USA
Nadia Zatsepin
Department of Physics, Arizona State University, Tempe, AZ 85287, USA
Uwe Weierstall
Department of Physics, Arizona State University, Tempe, AZ 85287, USA
Wei Liu
School of Molecular Sciences and Biodesign Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, AZ 85287, USA
Eriko Nango
RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan
Takanori Nakane
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 2-11-16 Yayoi, Bunkyo, Tokyo 113-0032, Japan
Rie Tanaka
RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan
Kensuke Tono
Japan Synchrotron Radiation Research Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198, Japan
Yasumasa Joti
Japan Synchrotron Radiation Research Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5198, Japan
So Iwata
RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan
Isabel Moraes
National Physical Laboratory, Hampton Road, Teddington TW11 0LW, England
Cornelius Gati
Department of Structural Biology, Stanford University, Stanford, CA 94305, USA
Vadim Cherezov
Bridge Institute, Departments of Chemistry and Biological Sciences, University of Southern California, Los Angeles, CA 90089, USA
Rational structure-based drug design (SBDD) relies on the availability of a large number of co-crystal structures to map the ligand-binding pocket of the target protein and use this information for lead-compound optimization via an iterative process. While SBDD has proven successful for many drug-discovery projects, its application to G protein-coupled receptors (GPCRs) has been limited owing to extreme difficulties with their crystallization. Here, a method is presented for the rapid determination of multiple co-crystal structures for a target GPCR in complex with various ligands, taking advantage of the serial femtosecond crystallography approach, which obviates the need for large crystals and requires only submilligram quantities of purified protein. The method was applied to the human β2-adrenergic receptor, resulting in eight room-temperature co-crystal structures with six different ligands, including previously unreported structures with carvedilol and propranolol. The generality of the proposed method was tested with three other receptors. This approach has the potential to enable SBDD for GPCRs and other difficult-to-crystallize membrane proteins.
