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. 2018 Aug 7;8(8):145. doi: 10.3390/brainsci8080145

Figure 3.

Figure 3

In silico modelling and in vitro functional analysis of the p.(Arg2His) mutation. (A) Ribbon models of alpha-tubulin (green) and beta-tubulin (blue) subunits aligned in a microtubule polymer. The position of Arg2 is shown (arrow) close to the inter-dimer interface (between alpha-tubulin and the beta-tubulin of an adjacent heterodimer). The mutation is on the opposite side of TUBA1A from the binding site of guanosine-5′-triphosphate (GTP, orange). (B) A close-up view of the Arg2 residue (arrow) with the mutant (purple ribbon, red side chain) and wild type (green ribbon and side chain) proteins superimposed. Only mild confirmation changes are predicted around the Arg2 residue. However, additional conformational changes are predicted between residues 38 and 51 (bracket). These may affect the interaction between heterodimers. (C) HEK-293 cells expressing FLAG-tagged TUBA1A-R2H. The cells are stained with DAPI (4′,6-diamidino-2-phenylindole, blue), anti-FLAG- (red), and anti-alpha-tubulin (green) antibodies. The microtubules appear yellow due to the colocalisation of endogenous (green) and FLAG-tagged transgenic (red) tubulin. The arrows indicate diffuse patches of transgenic mutant tubulin (red) in the cytoplasm between the microtubules. (D) Control cells expressing FLAG-tagged wild-type TUBA1A have less staining for the transgenic tubulin in the cytoplasm between the microtubules.