Regulation of Ca2+ channels by SNAP-25 via recruitment of syntaxin-1 from plasma membrane clusters
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Regulation of Ca2+ channels by SNAP-25 via recruitment of syntaxin-1 from plasma membrane clusters. / Toft-Bertelsen, Trine Lisberg; Ziomkiewicz, Iwona; Houy, Sébastien; da Silva Pinheiro, Paulo César; Sørensen, Jakob B.
In: Molecular Biology of the Cell, Vol. 27, No. 21, 01.11.2016, p. 3329-3341.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Regulation of Ca2+ channels by SNAP-25 via recruitment of syntaxin-1 from plasma membrane clusters
AU - Toft-Bertelsen, Trine Lisberg
AU - Ziomkiewicz, Iwona
AU - Houy, Sébastien
AU - da Silva Pinheiro, Paulo César
AU - Sørensen, Jakob B
N1 - © 2016 Toft-Bertelsen et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
PY - 2016/11/1
Y1 - 2016/11/1
N2 - SNAP-25 regulates Ca(2+) channels, with potentially important consequences for diseases involving an aberrant SNAP-25 expression level. How this regulation is executed mechanistically remains unknown. We investigated this question in mouse adrenal chromaffin cells and found that SNAP-25 inhibits Ca(2+) currents, with the B-isoform being more potent than the A-isoform, but not when syntaxin-1 is cleaved by botulinum neurotoxin C. In contrast, syntaxin-1 inhibits Ca(2+) currents independently of SNAP-25. Further experiments using immunostaining showed that endogenous or exogenous SNAP-25 expression recruits syntaxin-1 from clusters on the plasma membrane, thereby increasing the immunoavailability of syntaxin-1 and leading indirectly to Ca(2+) current inhibition. Expression of Munc18-1, which recruits syntaxin-1 within the exocytotic pathway, does not modulate Ca(2+) channels, whereas overexpression of the syntaxin-binding protein Doc2B or ubMunc13-2 increases syntaxin-1 immunoavailability and concomitantly down-regulates Ca(2+) currents. Similar findings were obtained upon chemical cholesterol depletion, leading directly to syntaxin-1 cluster dispersal and Ca(2+) current inhibition. We conclude that clustering of syntaxin-1 allows the cell to maintain a high syntaxin-1 expression level without compromising Ca(2+) influx, and recruitment of syntaxin-1 from clusters by SNAP-25 expression makes it available for regulating Ca(2+) channels. This mechanism potentially allows the cell to regulate Ca(2+) influx by expanding or contracting syntaxin-1 clusters.
AB - SNAP-25 regulates Ca(2+) channels, with potentially important consequences for diseases involving an aberrant SNAP-25 expression level. How this regulation is executed mechanistically remains unknown. We investigated this question in mouse adrenal chromaffin cells and found that SNAP-25 inhibits Ca(2+) currents, with the B-isoform being more potent than the A-isoform, but not when syntaxin-1 is cleaved by botulinum neurotoxin C. In contrast, syntaxin-1 inhibits Ca(2+) currents independently of SNAP-25. Further experiments using immunostaining showed that endogenous or exogenous SNAP-25 expression recruits syntaxin-1 from clusters on the plasma membrane, thereby increasing the immunoavailability of syntaxin-1 and leading indirectly to Ca(2+) current inhibition. Expression of Munc18-1, which recruits syntaxin-1 within the exocytotic pathway, does not modulate Ca(2+) channels, whereas overexpression of the syntaxin-binding protein Doc2B or ubMunc13-2 increases syntaxin-1 immunoavailability and concomitantly down-regulates Ca(2+) currents. Similar findings were obtained upon chemical cholesterol depletion, leading directly to syntaxin-1 cluster dispersal and Ca(2+) current inhibition. We conclude that clustering of syntaxin-1 allows the cell to maintain a high syntaxin-1 expression level without compromising Ca(2+) influx, and recruitment of syntaxin-1 from clusters by SNAP-25 expression makes it available for regulating Ca(2+) channels. This mechanism potentially allows the cell to regulate Ca(2+) influx by expanding or contracting syntaxin-1 clusters.
U2 - 10.1091/mbc.E16-03-0184
DO - 10.1091/mbc.E16-03-0184
M3 - Journal article
C2 - 27605709
VL - 27
SP - 3329
EP - 3341
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 21
ER -
ID: 169411819