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. 2008 Dec 31;3(12):e4074. doi: 10.1371/journal.pone.0004074

Figure 2. Munc18c binds directly to the cytosolic domains of Syntaxin 4 and VAMP2.

Figure 2

(A) 5 µg of either GST, or GST fused to the cytosolic domain of Sx4, VAMP2, VAMP 4 or VAMP 8, immobilised on glutathione Sepharose were incubated overnight at 4 C with 5 µg His-tagged Munc18c in a volume of 500 µl. After extensive washing in binding buffer, SDS-PAGE and immunoblot analysis was used to determine which of the GST proteins Munc18c had bound to. Upper panel represents a Coomassie stained gel of input proteins; lower panel shows an immunoblot for bound Munc18c (B) The ability of Munc18c to bind to a version of GST-VAMP2 harbouring only the SNARE motif of the v-SNARE was assessed as in (A). Data are representative of four experiments of this type.