ER-to-Golgi SNARE complex formation with Bos1 p.Arg196del.

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ER-to-Golgi SNARE complex formation with Bos1 p.Arg196del. ER-to-Golgi SNARE complex formation with Bos1 p.Arg196del. (A) SNARE complex formation was measured by the increase of fluorescence anisotropy as previously described (Demircioglu et al., 2014). For assembly, 500 nM OG-labeled Sec22 126-186C131OG were mixed with 2.7 µM Sed5 211-320, Bet1 1-118 and Bos1 151-221 or its mutants. A slower increase in fluorescence anisotropy indicates a reduced capability of Bos1 p.Arg196del to form the ER-to-Golgi SNARE complex. Bos1 p.Gly176Asp shows an even stronger effect. However, complex formation of Bos1 p.Gly176Trp seems to be accelerated. (B) Size exclusion chromatography (SEC) was performed for the individual SNARE domains of Bos1 variants. The Bos1 variants wild-type (wt), p.Gly176Asp and p.Arg196del seemed to be present in a higher oligomeric state than Bos1 p.Gly176Trp. Jörn M. Völker et al. Dis. Model. Mech. 2017;10:1391-1398 © 2017. Published by The Company of Biologists Ltd