Structural basis for recognition of synaptic vesicle protein 2C by botulinum neurotoxin A.
Botulinum neurotoxin A (BoNT/A) belongs to the most dangerous class of bioweapons. Despite this, BoNT/A is used to treat a wide range of common medical conditions such as migraines and a variety of ocular motility and movement disorders. BoNT/A is probably best known for its use as an antiwrinkle agent in cosmetic applications (including Botox and Dysport). BoNT/A application causes long-lasting flaccid paralysis of muscles through inhibiting the release of the neurotransmitter acetylcholine by cleaving synaptosomal-associated protein 25 (SNAP-25) within presynaptic nerve terminals. Two types of BoNT/A receptor have been identified, both of which are required for BoNT/A toxicity and are therefore likely to cooperate with each other: gangliosides and members of the synaptic vesicle glycoprotein 2 (SV2) family, which are putative transporter proteins that are predicted to have 12 transmembrane domains, associate with the receptor-binding domain of the toxin. Recently, fibroblast growth factor receptor 3 (FGFR3) has also been reported to be a potential BoNT/A receptor. In SV2 proteins, the BoNT/A-binding site has been mapped to the luminal domain, but the molecular details of the interaction between BoNT/A and SV2 are unknown. Here we determined the high-resolution crystal structure of the BoNT/A receptor-binding domain (BoNT/A-RBD) in complex with the SV2C luminal domain (SV2C-LD). SV2C-LD consists of a right-handed, quadrilateral β-helix that associates with BoNT/A-RBD mainly through backbone-to-backbone interactions at open β-strand edges, in a manner that resembles the inter-strand interactions in amyloid structures. Competition experiments identified a peptide that inhibits the formation of the complex. Our findings provide a strong platform for the development of novel antitoxin agents and for the rational design of BoNT/A variants with improved therapeutic properties.
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Roger M
RM
Benoit
1] Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland [2].
Daniel
D
Frey
1] Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland [2].
Manuel
M
Hilbert
1] Cell Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, The Netherlands [2].
Josta T
JT
Kevenaar
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Mara M
MM
Wieser
Swiss Light Source, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Christian U
CU
Stirnimann
UCB Celltech, UCB Pharma, UCB NewMedicines, Slough SL1 4EN, UK.
David
D
McMillan
UCB Celltech, UCB Pharma, UCB NewMedicines, Slough SL1 4EN, UK.
Tom
T
Ceska
UCB Pharma, UCB NewMedicines, B-1420 Braine-L'Alleud, Belgium.
Florence
F
Lebon
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Rolf
R
Jaussi
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Michel O
MO
Steinmetz
1] Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland [2] Department of Biology, ETH Zurich, CH-8093 Zurich, Switzerland.
Gebhard F X
GF
Schertler
Cell Biology, Faculty of Science, Utrecht University, 3584 CH Utrecht, The Netherlands.
Casper C
CC
Hoogenraad
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Guido
G
Capitani
Laboratory of Biomolecular Research, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland.
Richard A
RA
Kammerer
D001665:Binding Sites; D019274:Botulinum Toxins, Type A / Q000737:chemistry* / Q000378:metabolism*; D018360:Crystallography, X-Ray; D004705:Endocytosis / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D017072:Neostriatum / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship
D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; C585899:SV2C protein, human; D019274:Botulinum Toxins, Type A