Dominant antigenic peptides located at the heavy chain terminal of botulinum neurotoxin B contain receptor-binding sites for synaptotagmin II.
Botulism toxicity is caused by botulinum neurotoxins (BoNTs) that bind to the surface of the nerve terminals through the C-terminal portion of the heavy chain (Hc) and gain access to the neuron cells, thereby blocking the release of neurotransmitters into the synapse by the light chain. It has been recently found that synaptotagmin II (syt II) is the specific protein receptor of BoNTs; however, the molecular basis underlying the interaction between BoNTs and the receptor has been poorly understood. In this study, the recombinant fragment from the luminal domain of the human protein receptor syt II was prepared. Further, the Hc antigenic peptides of BoNTs type B (BoNTb) containing the possible epitopes were synthesized and analyzed for checking their ability for specific recognition and binding by using completely protective polyclonal antibody and were then evaluated for checking their ability to elicit protective immunity in mice against lethal toxin challenge. Finally, the interaction between the Hc-dominant antigenic peptides and syt II was analyzed. We found that two peptides located at the Hc terminal of the BoNTb showed the general character of the dominant antigenic epitopes in terms of protection against BoNTb intoxication. Further, they were found to contain the receptor-binding sites through the specific recognition of the receptor syt II. Both were derived from the uninterrupted segments of the amino acid residues H(1241-1277) of BoNTb. Kinetic data of the antigen-receptor interaction were depicted in real-time and calculated as affinity constant of 2.99x10(-7)M.
Shi
Jing
J
State Key Laboratory of Pathogens and Biosecurity, Institute of Microbiology and Epidemiology, No. 20 Dongdajie, Fengtai District, Beijing 100071, PR China.
Bao
Shizhong
S
Yin
Jun
J
Cai
Kun
K
Hou
Xiaojun
X
Xiao
Le
L
Tu
Wei
W
Wang
Qin
Q
Wang
Hui
H
D000595:Amino Acid Sequence; D000818:Animals; D001428:Bacterial Vaccines / Q000276:immunology; D001665:Binding Sites; D001905:Botulinum Toxins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D019274:Botulinum Toxins, Type A; D001906:Botulism / Q000276:immunology / Q000517:prevention & control; D005260:Female; D006801:Humans; D007114:Immunization; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D050861:Synaptotagmin II / Q000235:genetics / Q000378:metabolism*
D001428:Bacterial Vaccines; D016056:Immunodominant Epitopes; D010455:Peptides; D011994:Recombinant Proteins; C494262:SYT2 protein, human; D050861:Synaptotagmin II; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A