Author: David E. Gordon; Gwendolyn M. Jang; Mehdi Bouhaddou; Jiewei Xu; Kirsten Obernier; Matthew J O'Meara; Jeffrey Z. Guo; Danielle L. Swaney; Tia A. Tummino; Ruth Huttenhain; Robyn Kaake; Alicia L. Richards; Beril Tutuncuoglu; Helene Foussard; Jyoti Batra; Kelsey Haas; Maya Modak; Minkyu Kim; Paige Haas; Benjamin J. Polacco; Hannes Braberg; Jacqueline M. Fabius; Manon Eckhardt; Margaret Soucheray; Melanie Brewer; Merve Cakir; Michael J. McGregor; Qiongyu Li; Zun Zar Chi Naing; Yuan Zhou; Shiming Peng; Ilsa T. Kirby; James E. Melnyk; John S Chorba; Kevin Lou; Shizhong A. Dai; Wenqi Shen; Ying Shi; Ziyang Zhang; Inigo Barrio-Hernandez; Danish Memon; Claudia Hernandez-Armenta; Christopher J.P. Mathy; Tina Perica; Kala B. Pilla; Sai J. Ganesan; Daniel J. Saltzberg; Rakesh Ramachandran; Xi Liu; Sara B. Rosenthal; Lorenzo Calviello; Srivats Venkataramanan; Yizhu Lin; Stephanie A. Wankowicz; Markus Bohn; Phillip P. Sharp; Raphael Trenker; Janet M. Young; Devin A. Cavero; Joseph Hiatt; Theo Roth; Ujjwal Rathore; Advait Subramanian; Julia Noack; Mathieu Hubert; Ferdinand Roesch; Thomas Vallet; Björn Meyer; Kris M. White; Lisa Miorin; Oren S. Rosenberg; Kliment A. Verba; David Agard; Melanie Ott; Michael Emerman; Davide Ruggero; Adolfo Garcí-Sastre; Natalia Jura; Mark von Zastrow; Jack Taunton; Olivier Schwartz; Marco Vignuzzi; Christophe d'Enfert; Shaeri Mukherjee; Matt Jacobson; Harmit S. Malik; Danica G Fujimori; Trey Ideker; Charles S Craik; Stephen Floor; James S. Fraser; John Gross; Andrej Sali; Tanja Kortemme; Pedro Beltrao; Kevan Shokat; Brian K. Shoichet; Nevan J. Krogan
Title: A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug-Repurposing Document date: 2020_3_22
ID: 38d6gb7o_4
Snippet: Mature Nsps and all predicted proteins expressed from other SARS-CoV-2 Orfs (27 proteins plus one mutant) were codon optimized and cloned into a mammalian expression vector with a 2xStrep tag fused to either the Nor C-terminus 19 . Protein expression plasmids were transfected into human HEK293T cells, which were incubated for 40 hours before lysis in a mild detergent buffer. Viral proteins were affinity purified using MagStrep beads. Beads were w.....
Document: Mature Nsps and all predicted proteins expressed from other SARS-CoV-2 Orfs (27 proteins plus one mutant) were codon optimized and cloned into a mammalian expression vector with a 2xStrep tag fused to either the Nor C-terminus 19 . Protein expression plasmids were transfected into human HEK293T cells, which were incubated for 40 hours before lysis in a mild detergent buffer. Viral proteins were affinity purified using MagStrep beads. Beads were washed, on-bead digestion was performed overnight, and peptides were desalted and analyzed by protein mass spectrometry. High confidence interactors were identified using SAINTexpress and the MiST algorithm 19, 20 .
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