Abstract
The global pandemic of coronavirus disease 2019 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has become a severe global health problem because of its rapid spread. Both Ace2 and NRP1 provide initial viral binding sites for SARS-CoV-2. Here, we show that cysteine residues located in the vestigial plasminogen-apple-nematode (PAN) domain of NRP1 are necessary for SARS-CoV-2 spike protein internalization. Mutating novel cysteine residues in the PAN altered NRP1 stability and downstream activation of extracellular signal-regulated kinase (ERK) signaling pathway and impaired its interaction with the spike protein. This resulted in a significant reduction in spike protein abundance in Vero-E6 cells for the original, alpha, and delta SARS-CoV-2 variants even in the presence of the Ace2. Moreover, mutating these cysteine residues in NRP1 significantly lowered its association with Plexin-A1. As the spike protein is a critical component for targeted therapy, our biochemical study may represent a distinct mechanism to develop a path for future therapeutic discovery.
| Original language | English |
|---|---|
| Article number | 106274 |
| Journal | iScience |
| Volume | 26 |
| Issue number | 4 |
| DOIs | |
| State | Published - Apr 21 2023 |
Funding
Bioinformatics analyses of PAN domain distribution and functional inference was supported by the United States Department of Energy's Office of Science Early Career Research Program under the Biological and Environmental Research office. Biochemical, immunofluorescence, and transcriptome analyses in human cell lines were supported by the Oak Ridge National Laboratory Lab Directed Research Development program. Part of this research used resources at the Oak Ridge Leadership Computing Facility (OLCF) and the Compute and Data Environment for Science (CADES) at the Oak Ridge National Laboratory. Oak Ridge National Laboratory is managed by UT-Battelle, LLC for the U.S. Department of Energy under Contract Number DE-AC05-00OR22725. Debjani Pal was supported by the United States Department of Energy's Office of Science Lab Directed Research and Development fund under the Radioisotope Science and Technology Division Research office (grant number: 3130F004) during the revised version of manuscript preparation. The Vero-E6 cells were a generous gift from Carmen Foster at ORNL. Conceptualization: DP, KD, and WM. Methodology: DP, KD, TBY, and JK. Funding acquisition: WM. Writing – original draft: DP, KD, and WM. Writing – review & editing: DP, KD, TBY, and WM. The authors declare that they have no competing interests. Bioinformatics analyses of PAN domain distribution and functional inference was supported by the United States Department of Energy’s Office of Science Early Career Research Program under the Biological and Environmental Research office . Biochemical, immunofluorescence, and transcriptome analyses in human cell lines were supported by the Oak Ridge National Laboratory Lab Directed Research Development program . Part of this research used resources at the Oak Ridge Leadership Computing Facility (OLCF) and the Compute and Data Environment for Science (CADES) at the Oak Ridge National Laboratory. Oak Ridge National Laboratory is managed by UT-Battelle, LLC for the U.S. Department of Energy under Contract Number DE-AC05-00OR22725 . Debjani Pal was supported by the United States Department of Energy’s Office of Science Lab Directed Research and Development fund under the Radioisotope Science and Technology Division Research office (grant number: 3130F004 ) during the revised version of manuscript preparation. The Vero-E6 cells were a generous gift from Carmen Foster at ORNL.
Keywords
- Biochemistry
- Virology
