Structural and Functional Analysis of the CAPS SNARE-Binding Domain Required for SNARE Complex Formation and Exocytosis

Cell Rep. 2019 Mar 19;26(12):3347-3359.e6. doi: 10.1016/j.celrep.2019.02.064.

Abstract

Exocytosis of synaptic vesicles and dense-core vesicles requires both the Munc13 and CAPS (Ca2+-dependent activator proteins for secretion) proteins. CAPS contains a soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-binding region (called the DAMH domain), which has been found to be essential for SNARE-mediated exocytosis. Here we report a crystal structure of the CAPS-1 DAMH domain at 2.9-Å resolution and reveal a dual role of CAPS-1 in SNARE complex formation. CAPS-1 plays an inhibitory role dependent on binding of the DAMH domain to the MUN domain of Munc13-1, which hinders the ability of Munc13 to catalyze opening of syntaxin-1, inhibiting SNARE complex formation, and a chaperone role dependent on interaction of the DAMH domain with the syntaxin-1/SNAP-25 complex, which stabilizes the open conformation of Syx1, facilitating SNARE complex formation. Our results suggest that CAPS-1 facilitates SNARE complex formation via the DAMH domain in a manner dependent on sequential and cooperative interaction with Munc13-1 and SNARE proteins.

Keywords: CAPS; Munc13; SNARE; dense-core vesicle; exocytosis; synaptic vesicle.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Crystallography, X-Ray
  • HEK293 Cells
  • Humans
  • Nerve Tissue Proteins / chemistry*
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism
  • PC12 Cells
  • Protein Domains
  • Protein Structure, Quaternary
  • Rats
  • Sf9 Cells
  • Spodoptera
  • Synaptosomal-Associated Protein 25 / chemistry*
  • Synaptosomal-Associated Protein 25 / genetics
  • Synaptosomal-Associated Protein 25 / metabolism
  • Syntaxin 1 / chemistry*
  • Syntaxin 1 / genetics
  • Syntaxin 1 / metabolism

Substances

  • Nerve Tissue Proteins
  • SNAP25 protein, human
  • Synaptosomal-Associated Protein 25
  • Syntaxin 1
  • UNC13B protein, human