Regulated Secretion Is Impaired in AtT-20 Endocrine Cells Stably Transfected with Botulinum Neurotoxin Type A Light Chain

Fernando Aguado, Lourdes Gombau, Glòria Majó, Jordi Marsal, Jerónimo Blanco, Juan Blasi
1997 Journal of Biological Chemistry  
Botulinum neurotoxin type A (BoNT/A) inhibits neurotransmitter release by specific cleavage of SNAP-25, a synaptosome-associated protein also expressed in the ACTH secretory cell line AtT-20. Expression of light chain BoNT/A (L-BoNT/A) gene transfected into AtT-20 cells resulted in a cleaved form of SNAP-25 indistinguishable from that generated by bona fide BoNT/A. L-BoNT/A-transfected cells showed no difference in replication rate, viability, or phenotype, compared with control AtT-20 cells.
more » ... rol AtT-20 cells. In contrast, L-BoNT/A-transfected cells could not be induced to secrete ACTH upon stimulation by 8-bromo-cAMP or KCl. In addition, ␣-latrotoxin induced ACTH release from control cells, but not from L-BoNT/A-transfected cells. These experiments suggest an important role for SNAP-25 in regulated secretion from AtT-20 cells and underline the usefulness of this cell system as a tool for the study of the molecular mechanism of peptide hormone secretion.
doi:10.1074/jbc.272.41.26005 pmid:9325336 fatcat:fewjosthhjhbhcwqrooq3fm2zi