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

Interaction of syntaxin 1 with the alpha(1D) subunit of the voltage-gated L type Ca2+ channel was investigated in the pancreatic beta cell, Coexpressi on of the enhanced green fluorescent protein-linked alpha(1D) subunit with the enhanced blue fluorescent protein-linked syntaxin 1 and Western blot an alysis together with subcellular fractionation demonstrated that the alpha( 1D) subunit and syntaxin 1 were colocalized in the plasma membrane. Further more, the alpha(1D) subunit was coimmunoprecipitated efficiently by a polyc lonal 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 1 markedly reduced insulin release in b oth intact and permeabilized cells, subsequent to depolarization with K+ or exposure to high Ca2+, Exchanging Ca2+ for Ba2+ abolished the effect of an tisyntaxin 1 on both Ca2+ channel activity and insulin exocytosis, Moreover , antisyntaxin 1 had no significant effects on Ca2+-independent insulin rel ease trigged by hypertonic stimulation, This suggests that there is a struc ture-function relationship between the air, subunit of the L type Ca2+ chan nel and the exocytotic machinery in the pancreatic beta cell.