Tracking SNARE Complex Formation in Live Endocrine Cells

Seong J. An, and Wolfhard Almers^*

Abstract: Syntaxin, synaptosome-associated protein of 25 kD (SNAP25), and vesicle-associated membrane protein/synaptobrevin are collectively called SNAP receptor (SNARE) proteins, and they catalyze neuronal exocytosis by forming a "core complex." The steps in core complex formation are unknown. Here, we monitored SNARE complex formation in vivo with the use of a fluorescent version of SNAP25. In PC12 cells, we found evidence for a syntaxin-SNAP25 complex that formed with high affinity, required only the amino-terminal SNARE motif of SNAP25, tolerated a mutation that blocks formation of other syntaxin-SNAP25 complexes, and assembled reversibly when Ca²⁺ entered cells during depolarization. The complex may represent a precursor to the core complex formed during a Ca²⁺-dependent priming step of exocytosis.

Vollum Institute L-474, Oregon Health Sciences University, 3181 Southwest Sam Jackson Park Road, Portland, OR 97201, USA.

^* To whom correspondence should be addressed. E-mail: almersw{at}ohsu.edu

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