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Munc18 proteins bind non-cognate Sxs.

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posted on 31.10.2017, 17:47 authored by Michelle P. Christie, Shu-Hong Hu, Andrew E. Whitten, Asma Rehman, Russell J. Jarrott, Gordon J. King, Brett M. Collins, Jennifer L. Martin

Sx11-261-His or Sx41-275-His were incubated with detagged Munc18a or Munc18c for 2 h, 24 h or 48 h before the Sx was immobilized onto TALON Co2+ 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.