Fig. 6: Structure-guided mutagenesis at the HKU5:ACE2 interface.
From: ACE2 from Pipistrellus abramus bats is a receptor for HKU5 coronaviruses
a Co-structure for PDF2180 RBD and Pipistrellus pipistrellus ACE2 (PDB ID: 7wpz). b AlphaFold3 predicted structures for HKU5-20s RBD (residues 390-614) and Pipistrellus abramus ACE2 (residues 20-608). c AlphaFold3 predicted structure for HKU5-21s RBD (residues 390-609) and Pipistrellus abramus ACE2 (residues 20-608). d Comparison of viral RBD footprints on species ACE2. e Mutant spike expression in cell lysates and incorporation in pseudotyped particles. ACE2 residues common to all three interfaces are highlighted. BHK or 293 T cells transduced with either f Pipistrellus abramus (P.a.) or g human (H.s.) ACE2 were infected with indicated pseudotypes in quadruplicate as technical replicates. Dark circles indicate wildtype sequences, and triangles indicate point mutant sequences. For each graph, four individual entry values are plotted as points, the mean is shown as a bar and lines indicate standard deviation. 2-way ANOVA with Tukey correction for multiple comparisons (***) Indicates P value < 0.001. h Entry values for mutant RBDs as compared to parental wild-type RBD.
