Specific binding of human P[28] rotavirus VP8* protein to blood group ABH antigens on type 1 chains

Figures

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  Fig 1. Structural analysis of P[28]-VP8*. A. Crystal structure of P[28]-VP8* with two twisted antiparallel β-sheets. B. Superimposition of P[28]-VP8* structure to VP8*s of other genotypes. Values in the table represent root mean square deviations (RMSDs, unit: Å) of the Cα atoms of one VP8* monomer. VP8* structures were displayed in cartoon representation.

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  Fig 2. Glycan receptors of P[28]-VP8* detected by glycan microarray and the binding kinetics analysis. A. Binding signals of P[28]-VP8* on microarray. B. Binding signals of P[28]-VP8* pre-incubated with oligosaccharides at different concentration, ** P < 0.01, *** P < 0.001, **** P < 0.0001. C. BLI analysis of P[28]-VP8* with type 1 oligosaccharides.

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  Fig 3. Structural analysis of P[28]-VP8* in complex with H-T1 pentasaccharide LNFP I. A. Cartoon representation of the P[28]-VP8*-LNFP I complex. P[28]-VP8* was shown in ribbon representation with LNFP I shown in stick. The electron density maps of LNFP I contoured at 1σ level are shown at the right panel. B. Detailed interactions among the amino acids of VP8*-LNFP I with the hydrogen bonds represented by dotted lines. C. Schematic diagrams of the interactions. The details of the interaction between LNFP I and P[28]-VP8* were calculated with Ligplot. Hydrogen bonds are indicated by dashed lines between the atoms involved, while hydrophobic contacts are represented by arcs with spokes radiating toward the ligand atoms they contact. Atoms colored black, red, and blue represent carbon, oxygen, and nitrogen, respectively.

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  Fig 4. Determination of VP8* binding site and the key amino acids with blood group ABH on type 1 chains. A. On array inhibition of P[28]-VP8* binding to A-T1-Hexa and B-T1-Hexa using LNFP I as the inhibitor at different concentrations, *** P < 0.001, **** P < 0.0001. B. Molecular docking analysis of VP8* in complex with H-T1, A-T1, and B-T1 oligosaccharides. C. Molecular dynamics RMSD in complex with H-T1, A-T1, and B-T1 oligosaccharides. D. Microarray binding analysis of different P[28]-VP8* mutants in comparison with the wild-type, ns: P > 0.05, **** P < 0.0001.

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  Fig 5. Glycan binding properties of genetically closely-related RVs in the P[I] genogroup. A. Microarray binding of human P[10]-VP8*. B. Phylogenetic analysis of P[I] genogroup RVs (modified from Fig 1 of Liu et al 2016 [29]).

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  Fig 6. Comparison of the glycan binding site of P[28]-VP8* with other RV-VP8*. A. Comparation of the P[28] VP8*-LNFP I complex with published P[II] RV VP8* complexes. B. Surface representation of VP8*-LNFP I complex of P[28] and published VP8* complexes of RVs in the P[II] genogroup (the binding sites are in red). C. Comparison of the key amino acids of P[28]-VP8* in complex with LNFP I (pale green) with those of the P[8]-VP8* complex (deep salmon). D. Structure-based sequence alignment of VP8*s from selected P genotypes containing conserved amino acid residues (red) for binding to type 1 ABH (constructed using DNAMAN).