Journal: Journal of Biochemistry
Article Title: Electron microscopic imaging revealed the flexible filamentous structure of the cell attachment protein P2 of Rice dwarf virus located around the icosahedral 5-fold axes
doi: 10.1093/jb/mvv092
Figure Lengend Snippet: Overall structure of the isolated P2 protein. (a) SDS-PAGE of the structural proteins of RDV with and without CCl4 treatment. Intact RDV has seven kinds of proteins (Lane 2; P1–P3, P5, P7–P9 proteins). Lane 1, RDV particles lacking the P2 protein by CCl4 treatment used in the previous study (15); Lane 2, intact RDV particles containing the P2 protein (without CCl4 treatment) used in this study. P9 protein is appeared as several bands in SDS-PAGE (30). P9* is processed or degraded from a large polypeptide of P9 protein (P9). (b) SDS-PAGE and western blot analysis of the isolated P2 protein from intact RDV particles. The isolated P2 protein was separated by SDS-PAGE and then stained with Coomassie Brilliant Blue (left) or analysed by western blotting with RDV P2-specific antibodies (right). Lane 1, molecular weight markers; Lane 2, intact RDV particle; Lane 3, isolated P2 protein. (c) Electron micrograph of negatively stained P2 proteins isolated from intact RDV particles, which was recorded at a magnification of ×50,000 with an H7650 electron microscope (Hitachi, Japan). Small aggregated P2 proteins are also observed in the micrographs, which are indicated by arrowheads. The bar represents 50 nm. (d) Selected class-averaged images. The width of each image is 40.1 nm. A histogram of the lengths of the P2 proteins is presented later. The lengths of the P2 proteins were measured by tracing the centre of the filament in each class-averaged image. All 50 class-averaged images are shown in the Supplementary Data (Supplementary Fig. S2). (e) Schematic representation of the P2 structure.
Article Snippet: Abbreviations CCD charge-coupled device CPV Cytoplasmic polyhedrosis virus cryo-EM cryo-electron microscopy dsRNA double-stranded RNA RDV Rice dwarf virus SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis 3D three-dimensional
Techniques: Isolation, SDS Page, Western Blot, Staining, Molecular Weight, Microscopy
