Inhibition of botulinum neurotoxin a toxic action in vivo by synthetic synaptosome- and blocking antibody-binding regions.
In previous studies, we showed that certain peptides of the H(N) and H(C) domains of the H-chain of BoNT/A bind to mouse brain synaptosomes (snps). There was either complete correspondence or overlap between peptides that bind snps and those that bind human or mouse blocking antibodies (Abs). An equimolar mixture of the overlapping peptides N5/N6/N7/N8 (residues 505-523/519-537/533-551/547-565) extended the survival time of the mice to 74 h (20%) relative to controls, which had a 50% survival time of 60 h. On the other hand, peptide N26 (residues 799-817) provided no protection (50% survival time, 58 h), but the overlapping peptide N25 (785-803) almost doubled the 50% survival time to 119 h. A mixture of the overlap N25/N26 provided an unexpected level of protection permitting 40% of the mice to survive a lethal BoNT/A dose. In the H(C) domain, the overlap C23/C24 (1163-1181/1177-1195) provided no protection. Peptide C31 (1275-1296) also provided no significant protection. But an equimolar mixture of peptides C15/C16 (1051-1069/1065-1083) or peptides C18/C19/C20 (1093-1111/1107-1125/1121-1139) extended the 50% survival time by 41% (to 85 h) over controls (60 h) and was able to fully protect 20% of the mice which eventually recovered. Surprisingly, the mixture of the peptides C15/C16 and C18/C19/C20, which gave a 50% survival time of 75 h, was less protective than either peptides C15/C16 or peptides C18/C19/C20. The in vivo inhibitory activity of these peptides is discussed in relation to their location in the 3-dimensional structure of the toxin molecule and their membrane receptor binding.
Atassi
M Zouhair
MZ
Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX 77030, USA. matassi@bcm.edu
Dolimbek
Behzod Z
BZ
Steward
Lance E
LE
Aoki
K Roger
KR
D000595:Amino Acid Sequence; D000818:Animals; D019274:Botulinum Toxins, Type A / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D013574:Synaptosomes / Q000378:metabolism*
D010455:Peptides; D019274:Botulinum Toxins, Type A