Figure 2.

Formation of binary and ternary SNARE complexes with GST-Sft1p and GST-Sed5p. Soluble lysates from bacterial cells expressing recombinant GST- or (His)₆-tagged SNARE proteins (lacking their membrane anchor sequences) were mixed together and protein complexes isolated by affinity chromatography with glutathione Sepharose beads. Proteins were resolved by SDS-PAGE and stained with Coomassie brilliant blue. (A) GST-Sft1p binds to [Sed5p + Sec22p] (lane 1) and to [Sed5p] (lane 2), but binding to [Sec22p] is not detectable (lane 3). (B) GST-Sed5p binds [Sec2p + Bos1p] (lane 3) and [Sec22p] albeit weakly (lane 2) but not to [Bos1p] (lane 1). The star symbol (★) in Figure 2A denotes GST derived from proteolysis of GST-Sft1p. The dots either side of lane 2 in A highlight the presence of Sed5p.