Dimer Organization of Membrane?Associated NS5A of Hepatitis C Virus as Determined by Highly Sensitive ¹ H?Detected Solid?State NMR

The Hepatitis C virus nonstructural protein 5A (NS5A) is a membrane-associated involved in multiple steps of the viral life cycle. Direct-acting antivirals (DAAs) targeting NS5A are cornerstone antiviral therapy, but mode-of-action these drugs poorly understood. This due to lack information on membrane-bound structure. Herein, we present structural model an AH-linker-D1 reconstituted as proteoliposomes. We use highly sensitive proton-detected solid-state NMR methods suitable study samples generated through synthetic biology approaches. Spectra analyses disclose that both AH membrane anchor and linker flexible. Paramagnetic relaxation enhancements (PRE) reveal dimer organization lipids requires new type self-interaction not reflected previous crystal structures. In conclusion, provide first characterization lipidic environment shedding light onto clinically used inhibitors.