10.1371/journal.pone.0187302.g002 Michelle P. Christie Michelle P. Christie Shu-Hong Hu Shu-Hong Hu Andrew E. Whitten Andrew E. Whitten Asma Rehman Asma Rehman Russell J. Jarrott Russell J. Jarrott Gordon J. King Gordon J. King Brett M. Collins Brett M. Collins Jennifer L. Martin Jennifer L. Martin Munc18 proteins bind non-cognate Sxs. Public Library of Science 2017 non-cognate Munc 18c Syntaxin 1 Syntaxin 4. Munc 18a Neuronal Munc 18a SNARE protein Syntaxin Revisiting interaction specificity SNARE interaction specificity Syntaxin N-peptide interaction Munc 18c Munc 18a Regulatory Munc 18 proteins SNARE proteins partner SNARE Syntaxins spatiotemporal fusion events Munc 18c bind Syntaxins SNARE partners SNAP 25 binding Syntaxin 4 binding interactions adipocyte Munc 18 membrane fusion Syntaxin 1 N-peptide VAMP Syntaxin 1 Syntaxin 4 adipocyte Munc 18c 2017-10-31 17:47:43 Figure https://plos.figshare.com/articles/figure/Munc18_proteins_bind_non-cognate_Sxs_/5557249 <p>Sx1<sub>1-261</sub>-His or Sx4<sub>1-275</sub>-His were incubated with detagged Munc18a or Munc18c for 2 h, 24 h or 48 h before the Sx was immobilized onto TALON<sup>™</sup> Co<sup>2+</sup> affinity beads for 1h and then washed. Samples of the beads were then run on SDS-PAGE and stained with Coomassie Blue to determine if detagged Munc18 had been pulled down by cognate and non-cognate Sx partners. Detagged Munc18a and Munc18c were also incubated for the same time periods on beads without bound Sx to monitor non-specific binding (control lanes). Solid vertical lines on the gel image denote the removal of intervening lanes or placing two different gels adjacent to each other.</p>