Case Study of High-Throughput Drug Screening and Remote Data Collection for SARS-CoV-2 Main Protease by Using Serial Femtosecond X-ray Crystallography
Omur Guven,
Mehmet Gul,
Esra Ayan,
J Austin Johnson,
Baris Cakilkaya,
Gozde Usta,
Fatma Betul Ertem,
Nurettin Tokay,
Busra Yuksel,
Oktay Gocenler,
Cengizhan Buyukdag,
Sabine Botha,
Gihan Ketawala,
Zhen Su,
Brandon Hayes,
Frederic Poitevin,
Alexander Batyuk,
Chun Hong Yoon,
Christopher Kupitz,
Serdar Durdagi,
Raymond G. Sierra,
Hasan DeMirci
Affiliations
Omur Guven
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Mehmet Gul
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Esra Ayan
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
J Austin Johnson
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Baris Cakilkaya
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Gozde Usta
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Fatma Betul Ertem
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Nurettin Tokay
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Busra Yuksel
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Oktay Gocenler
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Cengizhan Buyukdag
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Sabine Botha
Department of Physics, Arizona State University, Tempe, AZ 85287, USA
Gihan Ketawala
Department of Physics, Arizona State University, Tempe, AZ 85287, USA
Zhen Su
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Brandon Hayes
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Frederic Poitevin
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Alexander Batyuk
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Chun Hong Yoon
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Christopher Kupitz
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Serdar Durdagi
Computational Biology and Molecular Simulations Laboratory, Department of Biophysics, School of Medicine, Bahcesehir University, Istanbul 34734, Turkey
Raymond G. Sierra
Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Rd., Menlo Park, CA 94025, USA
Hasan DeMirci
Department of Molecular Biology and Genetics, Koc University, Istanbul 34450, Turkey
Since early 2020, COVID-19 has grown to affect the lives of billions globally. A worldwide investigation has been ongoing for characterizing the virus and also for finding an effective drug and developing vaccines. As time has been of the essence, a crucial part of this research has been drug repurposing; therefore, confirmation of in silico drug screening studies have been carried out for this purpose. Here we demonstrated the possibility of screening a variety of drugs efficiently by leveraging a high data collection rate of 120 images/second with the new low-noise, high dynamic range ePix10k2M Pixel Array Detector installed at the Macromolecular Femtosecond Crystallography (MFX) instrument at the Linac Coherent Light Source (LCLS). The X-ray Free-Electron Laser (XFEL) is used for remote high-throughput data collection for drug repurposing of the main protease (Mpro) of SARS-CoV-2 at ambient temperature with mitigated X-ray radiation damage. We obtained multiple structures soaked with nine drug candidate molecules in two crystal forms. Although our drug binding attempts failed, we successfully established a high-throughput Serial Femtosecond X-ray crystallographic (SFX) data collection protocol.
