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In vitro translation of type A Clostridium botulinum neurotoxin heavy chain and analysis of its binding to rat synaptosomes.

作者信息

Li L, Singh B R

机构信息

Department of Chemistry and Biochemistry, University of Massachusetts Dartmouth, 02747, USA.

出版信息

J Protein Chem. 1999 Jan;18(1):89-95. doi: 10.1023/a:1020655701852.

DOI:10.1023/a:1020655701852
PMID:10071933
Abstract

Botulinum neurotoxins (BoNTs) are highly potent toxins that inhibit neurotransmitter release from peripheral cholinergic synapses. BoNTs consist of a toxifying light chain (LC; 50 kDa) and a binding/translocating heavy chain (HC; 100 kDa) linked through a disulfide bond. A DNA fragment encoding type A Clostridium botulinum heavy chain (BoNT/A HC) was amplified by polymerase chain reaction and cloned into an E. coli PET-15b vector. In vitro translated [35S]BoNT/A HC was identified by anti-BoNT/A polyclonal antibodies, and was used to investigate the binding of the toxin to rat synaptosomes. The binding of [35S]BoNT/A HC to synaptosomes was abolished by 500-fold excess of cold BoNT/A, and by incubation with trypsin. Treatment of BoNT/A HC with anti-BoNT/A or G(T1b) blocked its binding to synaptosomes. The radioactive BoNT/A HC recognized three proteins corresponding to a molecular mass of 150 (P150), 120 (P120), and 75 (P75) kDa in rat and bovine synaptosomal preparations. These results represent the first successful expression of functional full-length BoNT heavy chain.

摘要

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引用本文的文献

1
The C-terminal heavy-chain domain of botulinum neurotoxin a is not the only site that binds neurons, as the N-terminal heavy-chain domain also plays a very active role in toxin-cell binding and interactions.肉毒杆菌神经毒素A的C末端重链结构域并非唯一与神经元结合的位点，因为N末端重链结构域在毒素与细胞的结合及相互作用中也发挥着非常积极的作用。
Infect Immun. 2015 Apr;83(4):1465-76. doi: 10.1128/IAI.00063-15. Epub 2015 Jan 26.
2
Synaptotagmin II and gangliosides bind independently with botulinum neurotoxin B but each restrains the other.突触融合蛋白 II 和神经节苷脂与肉毒杆菌神经毒素 B 独立结合，但两者彼此抑制。
Protein J. 2014 Jun;33(3):278-88. doi: 10.1007/s10930-014-9557-y.
3

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Ganglioside GD3 enhances adherence of botulinum and tetanus neurotoxins to bovine brain synapsin I.神经节苷脂GD3增强肉毒杆菌和破伤风神经毒素对牛脑突触素I的黏附。
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