Syntaxin 1A drives fusion of large dense-core neurosecretory granules into a planar lipid bilayer

Cell Biochemistry and Biophysics
James M McNallyJosé R Lemos

Abstract

The SNARE complex, involved in vesicular trafficking and exocytosis, is composed of proteins in the vesicular membrane (v-SNAREs) that intertwine with proteins of the target membrane (t-SNAREs). Our results show that modified large dense-core neurosecretory granules (NSGs), isolated from the bovine neurohypophysis, spontaneously fuse with a planar lipid membrane containing only the t-SNARE syntaxin 1A. This provides evidence that syntaxin alone is able to form a functional fusion complex with native v-SNAREs of the NSG. The fusion was similar to constitutive, not regulated, exocytosis because changes in free [Ca2+] had no effect on the syntaxin-mediated fusion. Several deletion mutants of syntaxin 1A were also tested. The removal of the regulatory domain did not significantly reduce spontaneous fusion. However, a syntaxin deletion mutant consisting of only the transmembrane domain was incapable of eliciting spontaneous fusion. Finally, a soluble form of syntaxin 1A (lacking its transmembrane domain) was used to saturate the free syntaxin-binding sites of modified NSGs. This treatment blocks spontaneous fusion of these granules to a bilayer containing full-length syntaxin 1A. This method provides an effective model system to stu...Continue Reading

Citations

May 28, 2014·The Journal of General Physiology·James M McNallyJosé R Lemos
May 20, 2006·Proceedings of the National Academy of Sciences of the United States of America·Mark Bowen, Axel T Brunger
Dec 4, 2012·Chemistry and Physics of Lipids·David E LeeDixon J Woodbury
Aug 5, 2008·Trends in Neurosciences·Vladimir Parpura, U Mohideen
Jun 22, 2010·Journal of Neurochemistry·Inmaculada López-FontLuis M Gutiérrez
May 2, 2006·Biophysical Journal·W LiuVladimir Parpura
Aug 2, 2005·Biophysical Journal·Tingting LiuJames C Weisshaar
Sep 28, 2016·Journal of Medicinal Chemistry·Rongjun HeZhong-Yin Zhang
Jan 12, 2017·Biophysical Journal·Jason PaxmanDixon J Woodbury
Aug 31, 2019·Nature Communications·Alex J B KreutzbergerLukas K Tamm

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