Syntaxin 1 interacts with the L(D) subtype of voltage-gated Ca2+ channels in pancreatic β cells

S. N. Yang, O. Larsson, R. Branstrom, A. M. Bertorello, B. Leibiger, I. B. Leibiger, T. Moede, M. Kohler, B. Meister, P. O. Berggren

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Abstract

Interaction of syntaxin 1 with the α(1D) subunit of the voltage-gated L type Ca2+ channel was investigated in the pancreatic β cell. Coexpression of the enhanced green fluorescent protein-linked α(1D) subunit with the enhanced blue fluorescent protein-linked syntaxin I and Western blot analysis together with subcellular fractionation demonstrated that the α(1D) subunit and syntaxin 1 were colocalized in the plasma membrane. Furthermore, the α(1D) subunit was coimmunoprecipitated efficiently by a polyclonal antibody against syntaxin 1. Syntaxin 1 also played a central role in the modulation of L type Ca2+ channel activity because there was a faster Ca2+ current run-down in cells incubated with antisyntaxin 1 compared with controls. In parallel, antisyntaxin I markedly reduced insulin release in both intact and permeabilized cells, subsequent to depolarization with K+ or exposure to high Ca2+. Exchanging Ca2+ for Ba2+ abolished the effect of antisyntaxin 1 on both Ca2+ channel activity and insulin exocytosis. Moreover, antisyntaxin 1 had no significant effects on Ca2+-independent insulin release trigged by hypertonic stimulation. This suggests that there is a structure-function relationship between the α(1D) subunit of the L type Ca2+ channel and the exocytotic machinery in the pancreatic β cell.

Original languageEnglish
Pages (from-to)10164-10169
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume96
Issue number18
DOIs
StatePublished - Aug 31 1999
Externally publishedYes

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ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Yang, S. N., Larsson, O., Branstrom, R., Bertorello, A. M., Leibiger, B., Leibiger, I. B., Moede, T., Kohler, M., Meister, B., & Berggren, P. O. (1999). Syntaxin 1 interacts with the L(D) subtype of voltage-gated Ca2+ channels in pancreatic β cells. Proceedings of the National Academy of Sciences of the United States of America, 96(18), 10164-10169. https://doi.org/10.1073/pnas.96.18.10164