Plasma Membrane Localization Signals in the Light Chain of Botulinum Neurotoxin

Botulinum neurotoxin (BoNT) is a potent biological substance used to treat neuromuscular and pain disorders. Both BoNT type A and BoNT type E display high-affinity uptake into motor neurons and inhibit exocytosis through cleavage of the synaptosome-associated protein of 25 kDa (SNAP25). The therapeu...

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Published in:	Proceedings of the National Academy of Sciences - PNAS Vol. 101; no. 9; pp. 3208 - 3213
Main Authors:	Fernández-Salas, Ester, Steward, Lance E., Ho, Helen, Garay, Patton E., Sun, Sarah W., Gilmore, Marcella A., Ordas, Joseph V., Wang, Joanne, Francis, Joseph, Aoki, K. Roger, Boyer, Herbert W.
Format:	Journal Article
Language:	English
Published:	United States National Academy of Sciences 02-03-2004 National Acad Sciences
Subjects:	Amino Acid Sequence Animals Antibodies Binding Sites Biological Sciences Blotting, Western Botulinum Toxins - chemistry Botulinum Toxins - metabolism Botulinum Toxins, Type A - chemistry Botulinum Toxins, Type A - metabolism Carrier Proteins - metabolism Cell Membrane - physiology Cell membranes Cells Cellular differentiation Chromosomal Proteins, Non-Histone Clostridium botulinum Cytoplasm Exocytosis Fluorescence Leucine Light Macromolecular Substances Membranes Nerve Tissue Proteins - metabolism Neurology Neurons Neurotoxins PC12 Cells Plasma Proteins Rats Recombinant Proteins - metabolism Toxins Transfection
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Summary:	Botulinum neurotoxin (BoNT) is a potent biological substance used to treat neuromuscular and pain disorders. Both BoNT type A and BoNT type E display high-affinity uptake into motor neurons and inhibit exocytosis through cleavage of the synaptosome-associated protein of 25 kDa (SNAP25). The therapeutic effects of BoNT/A last from 3 to 12 months, whereas the effects of BoNT/E last less than 4 weeks. Using confocal microscopy and site-specific mutagenesis, we have determined that the protease domain of BoNT/A light chain (BoNT/A-LC) localizes in a punctate manner to the plasma membrane, colocalizing with the cleaved product, SNAP25197. In contrast, the short-duration BoNT/E serotype is cytoplasmic. Mutations in the BoNT/A-LC have revealed sequences at the N terminus necessary for plasma membrane localization, and an active dileucine motif in the C terminus that is likely involved in trafficking and interaction with adaptor proteins. These data support sequence-specific signals as determinants of intracellular localization and as a basis for the different durations of action in these two BoNT serotypes.
Bibliography:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 To whom correspondence may be addressed. E-mail: fernandez_ester@allergan.com or aoki_roger@allergan.com. Communicated by Herbert W. Boyer, University of California, San Francisco, CA, January 12, 2004 Abbreviations: BoNT, botulinum neurotoxin; LC, light chain; SNAP25, synaptosome-associated protein of 25 kDa.
ISSN:	0027-8424 1091-6490
DOI:	10.1073/pnas.0400229101