Regulation of syntaxin1A-munc18 complex for SNARE pairing in HEK293 cells.
Svetlana E Gladycheva, Chi S Ho, Yue Ying F Lee, Edward L Stuenkel
Author Information
Svetlana E Gladycheva: 7804 Medical Sciences II Building, Department of Molecular and Integrative Physiology, The Medical School, University of Michigan, Ann Arbor, MI 48109-0622, USA. segladys@umich.edu
The formation and dissolution of SNARE protein complexes is essential for Ca(2+)-triggered fusion of neurotransmitter-filled vesicles at the presynaptic membrane. Among the pre-synaptic SNARE proteins, the activation of the Q-SNAREsyntaxin1A is a critical event for SNARE complex formation. Activation requires syntaxin1A to transit from a munc18-bound non-interacting state to one competent for SNARE binding. The molecular mechanisms that regulate this transition remain unclear. The propensity of syntaxin1A to promote voltage-dependent steady-state inactivation of N-type Ca(2+) channels and accelerate their entry into inactivation was used in a heterologous cell expression system to elucidate regulation of syntaxin1A protein-protein interactions. We report that coexpression of munc18 eliminated the promoting effect of syntaxin1A on inactivation. This effect of munc18 was completely disrupted by coexpression of munc13-1, but not munc13-2 or munc13-3. Also, since expression of munc13-1 with syntaxin1A resulted in an inactivation phenotype identical to that of munc18 with syntaxin1A, the action of munc13-1 on the munc18-syntaxin1A complex was functionally unique and did not result from competitive binding interactions. Furthermore, munc13 expressed with syntaxin1A and munc18 promoted redistribution of a cytosolic SNAP25 mutant to the membrane, a result indicative of syntaxin1A-SNAP25SNARE pairing. These data demonstrate an important role of munc13 to control the protein-protein interactions of syntaxin1A in vivo, and support munc13 as critical to dissociating syntaxin1A-munc18 complexes and making syntaxin1A available for SNARE interactions.
References
Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7363-8
[PMID: 8692999]
