Mode of VAMP substrate recognition and inhibition of Clostridium botulinum neurotoxin F.
Clostridium botulinum neurotoxins (BoNTs) cleave neuronal proteins responsible for neurotransmitter release, causing the neuroparalytic disease botulism. BoNT serotypes B, D, F and G cleave and inactivate vesicle-associated membrane protein (VAMP), each at a unique peptide bond. The specificity of BoNTs depends on the mode of substrate recognition. We have investigated the mechanism of substrate recognition of BoNT F by determining the crystal structures of its complex with two substrate-based inhibitors, VAMP 22-58/Gln58D-cysteine and 27-58/Gln58D-cysteine. The inhibitors bind to BoNT F in the canonical direction (as seen for BoNTs A and E substrates) but are positioned specifically via three major exosites away from the active site. The cysteine sulfur of the inhibitors interacts with the zinc and exists as sulfinic acid in the inhibitor VAMP 27-58/Gln58D-cysteine. Arg133 and Arg171, which form part of two separate exosites, are crucial for substrate binding and catalysis.
Agarwal
Rakhi
R
Biology Department, Brookhaven National Laboratory, Upton, New York, USA.
Schmidt
James J
JJ
Stafford
Robert G
RG
Swaminathan
Subramanyam
S
D000595:Amino Acid Sequence; D001905:Botulinum Toxins / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D003545:Cysteine / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism*; D050683:R-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D013455:Sulfur / Q000737:chemistry
D020033:Protein Isoforms; D050683:R-SNARE Proteins; D013455:Sulfur; D001905:Botulinum Toxins; D003545:Cysteine; C096321:botulinum toxin type F