Stu2p: A microtubule-binding protein that is an essential component of the yeast spindle pole body. J Cell Biol 139, 1271–1280 (1997). 51. Al-Bassam, J. & Chang, F. Regulation of microtubule dynamics by TOG-domain proteins XMAP215/Dis1 and CLASP. Trends Cell Biol 21, 604–614 (...
aSchematic representation of the domain structure of recombinant ch-TOG and truncation mutants carrying a C-terminal GFP tag. TOG domains colored in yellow, C-terminal domain in cyan.bU2OS wild type cells or cells stably expressing recombinant GFP-tagged ch-TOG constructs as in (a) were transfe...
However, the internal loop is predicted to form a β sheet structure, implying that this domain might represent a protein-protein interaction motif [6, 7]. To address this point, we examined whether Ndc80 and Dis1 physically interact. Coimmunoprecipitation experiments showed that it is the case...
Fractions containing the target protein were pooled and loaded onto 500 μ l of Ni-NTA Superflow resin (Qiagen) equilibrated with low-imidazole buffer. The resin was washed with 10 ml of low-imidazole buffer then imidazole buffer containing 30 and 60 mM imidazole buffer before elution...