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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Biol+Chem
2014 ; 289
(24
): 17087-99
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Differential interaction of tomosyn with syntaxin and SNAP25 depends on domains
in the WD40 ?-propeller core and determines its inhibitory activity
#MMPMID24782308
Bielopolski N
; Lam AD
; Bar-On D
; Sauer M
; Stuenkel EL
; Ashery U
J Biol Chem
2014[Jun]; 289
(24
): 17087-99
PMID24782308
show ga
Neuronal exocytosis depends on efficient formation of soluble
N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes
and is regulated by tomosyn, a SNARE-binding protein. To gain new information
about tomosyn's activity, we characterized its mobility and organization on the
plasma membrane (PM) in relation to other SNARE proteins and inhibition of
exocytosis. By using direct stochastic optical reconstruction microscopy
(dSTORM), we found tomosyn to be organized in small clusters adjacent to syntaxin
clusters. In addition, we show that tomosyn is present in both syntaxin-tomosyn
complexes and syntaxin-SNAP25-tomosyn complexes. Tomosyn mutants that lack
residues 537-578 or 897-917 from its ?-propeller core diffused faster on the PM
and exhibited reduced binding to SNAP25, suggesting that these mutants shift the
equilibrium between tomosyn-syntaxin-SNAP25 complexes on the PM to
tomosyn-syntaxin complexes. As these deletion mutants impose less inhibition on
exocytosis, we suggest that tomosyn inhibition is mediated via
tomosyn-syntaxin-SNAP25 complexes and not tomosyn-syntaxin complexes. These
findings characterize, for the first time, tomosyn's dynamics at the PM and its
relation to its inhibition of exocytosis.