Minor electrostatic changes robustly increase VP40 membrane binding, assembly, and budding of Ebola virus matrix protein derived virus-like particles

Development of a Crystallographic Screening to Identify Sudan Virus VP40 Ligands

“…Notably, the mutations markedly reduced the efficiency of budding, with the most pronounced decrease observed for the sVP40 L213A mutant (Figure B,C). These findings are consistent with the existing literature; specifically, residues L213 and R214 have been previously investigated in the context of eVP40. , A recent study highlighted the importance of electrostatics of the linker residues between the NTD and CTD for the interaction with the cellular plasma membrane and showed that differences in the net charge of VP40 alter budding dynamics . To investigate a similar mechanism for the pocket residues, we prepared various R214 mutants and assessed their functionality in an MG and budding assay (Figure D,E) and could show that the net charge plays only a minor role in the regulation of viral RNA synthesis (especially when compared to R214A, Figure A).…”

“… 39 , 40 A recent study highlighted the importance of electrostatics of the linker residues between the NTD and CTD for the interaction with the cellular plasma membrane and showed that differences in the net charge of VP40 alter budding dynamics. 41 To investigate a similar mechanism for the pocket residues, we prepared various R214 mutants and assessed their functionality in an MG and budding assay ( Figure 3 D,E) and could show that the net charge plays only a minor role in the regulation of viral RNA synthesis (especially when compared to R214A, Figure 3 A). In contrast, defects in budding activity are more prominent for negative or uncharged substitutions compared to other substitutions that retain net charges, such as R214K or R214H ( Figure 3 E).…”