N-glycoproteomic profiling reveals new therapeutic targets against coronavirus infestation which may be involved in Cepharanthine’s intervention

ABSTRACT: N-glycosylation is an important post-translational modification involved in protein folding, signal transduction, extracellular matrix (ECM) organization and immune response. Recent evidence shows that SARS-CoV-2 Spike protein is highly glycosylated and it may be potential target in viral pathology and drug/vaccine design. Therefore, the N-glycoproteomic profiling of coronavirus and its infected cells are of great importance in therapeutic targets screening for drug discovery. Here, we carried out 4D label-free LC-MS/MS-based N-glycoproteomics using a well-established SARS-CoV-2 cellular model, pangolin GX_P2V virus-infected Vero E6 cells, to study the mechanism of coronavirus infestation and potential drug targets. Meanwhile, we investigated the effect of Cepharanthine (CEP) on viral-induced aberrant N-glycoprotein changes in affected cells and on the viral proteins. The results showed that coronavirus GX_P2V could cause aberrant glycosylation of cell proteins at multiple levels, including extracellular matrix (ECM) and related signal transduction, whereas CEP can maintain 12 out of 69 GX_P2V-induced aberrant glycoproteins at normal glycosylation state. Functional enrichment and PPI analyses revealed that LAMB1 and FN1 were the pivotal proteins in regulating the aberrant glycosylation caused by coronavirus in presence of CEP, indicating that CEP might achieve its therapeutic intervention via these potential targets. Besides, CEP can regulate the glycosylation of viral proteins S, M and N. Nevertheless, there were still 57 out of 69 glycoproteins which cannot be significantly affected by CEP, indicating the combination of CEP with other drugs against the rest of targets should be considered.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Chlorocebus Sabaeus

TISSUE(S): Cell Culture

SUBMITTER: Wenlin An  

LAB HEAD: Wenlin An

PROVIDER: PXD021297 | Pride | 2022-08-12

REPOSITORIES: Pride