SNAP-25 is present in a SNARE complex in adrenal chromaffin cells

Abstract

SNAP-25 (synaptosomal-associated protein 25 kDa) is a target for botulinum neurotoxins A and E, which both inhibit neurotransmitter release, and was recently identified together with syntaxin and synaptobrevin as receptors for NSF and alpha-SNAP. We show that SNAP-25 was enriched in the microsomal fraction from adrenal medulla, although the level of SNAP-25 in adrenal medullary microsomes was about 20-fold less than in brain microsomes. Immunocytochemistry confirmed the presence of SNAP-25 in cultured chromaffin cells and showed plasma membrane staining. Using immunoprecipitation, we found that SNAP-25 was present in a complex with syntaxin, synaptobrevin, synaptotagmin, NSF, alpha-SNAP and other unidentified polypeptides. These data indicate that SNAP-25 in chromaffin cells is present in a complex similar to that identified in brain.