Fig. 1: Biochemical and structural characterization of A. thaliana Augmin assemblies.
A Schematic representation of plant Augmin hetero-octamer (AUG1,2,3,4,5,6,7,8) subunit organization, showing domain boundaries and purification tags. Note: AUG8 construct (residues 383–644) excludes the N-terminal MT binding domain (See Supplementary Fig. 1). B Biochemical validation of hetero-octameric Augmin complex. Left: SEC-MALS analysis confirming monodisperse assembly with predicted octameric mass (single run). Right: SDS-PAGE of purified complex showing all eight subunits (See Supplementary Fig. 1). AUG6* marks a degradation product of AUG6. C Schematic of minimal Augmin hetero-tetramer (AUG1,3,4,5) subunit organization, including domain boundaries and purification tags. D Biochemical validation of hetero-tetrameric complex. Left: SEC-MALS analysis demonstrating monodisperse assembly with tetrameric mass (Single run). Right: SDS-PAGE confirming presence of four subunits. E Cryo-EM 2D-class averages revealing Augmin complex architecture. Top panel: Monomeric AUG1,2,3,4,5,6,7,8: 40 nm tuning fork structure. Second panel: Monomeric AUG1,2,3,4,5,6,7,8: V-junction focus with extended region. Third panel: Dimeric AUG1,2,3,4,5,6,7,8: Two extended regions, single V-junction. Fourth panel: Dimeric AUG1,2,3,4,5,6,7,8: Two extended regions, two V-junctions. Fifth panel: Focused V-junction from AUG1,2,3,4,5,6,7,8: 24-nm V-junction and stem regions. Bottom Panel: Monomeric AUG1,3,4,5: 23 nm extended region.
