Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry.
Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry.
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通过基质辅助激光解吸电离和电喷雾质谱法表征 C、D、E 和 F 型肉毒杆菌毒素。
DOI：
10.1016/j.chroma.2004.02.047
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
E. Wils
中科院分区：
文献类型：
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作者：
B. V. van Baar;A. Hulst;A. D. de Jong;E. Wils
在通过质谱（MS）对a型和B型肉毒毒素（肉毒毒素a和肉毒毒素B）的早期特征进行随访后，现在研究了C型，D型，E型和F型（肉毒毒素C，肉毒毒素D，肉毒毒素E，肉毒毒素F）。肉毒杆菌毒素是一种极具神经毒性的细菌毒素，可能被用作生物战剂。具有生物活性的肉毒毒素c、肉毒毒素d、肉毒毒素e和肉毒毒素f是由各自的神经毒素与无毒的非血凝素（NTNH）和有时是特异性血凝素（HA）的蛋白质复合物组成的。这些蛋白复合物在质谱鉴定中被观察到。肉毒梭菌003-9的BTxC复合体由一种类似于C型菌株6813的“C1型和D型镶嵌”毒素、一种类似于C- stockholm菌株的无毒无血凝成分和一种类似于C- stockholm菌株的33kDa血凝成分（HA-33）和一种外酶C3组成，其序列与菌株003-9的已知基因序列完全一致。肉毒杆菌CB-16的BTxD复合物含有与D型菌株BVD/-3相同的神经毒素和与C型菌株C- yoichi相同的NTNH。值得注意的是，观察到的CB-16 NTNH蛋白序列与已知基因序列L859而不是F859相差一个氨基酸。从鲱鱼中分离的肉毒杆菌BTxE复合体，其神经毒素序列与菌株NCTC 11219相同，NTNH与E型菌株Mashike相似（与观察序列相差1个氨基酸）。BTxF来源于肉毒杆菌Langeland菌株（NCTC 10281），由神经毒素和NTNH组成；观察到的两种蛋白序列与已知菌株Langeland的基因序列一致。与BTxA和BTxB一样，基质辅助激光解吸/电离（MALDI）质谱（MS）从胰蛋白酶消化肽图中提供临时鉴定，液相色谱-电喷雾（串联）质谱（LC-ES MS）从胰蛋白酶消化获得的消化肽的氨基酸序列信息中提供明确鉴定。
In a follow-up of the earlier characterisation of botulinum toxins type A and B (BTxA and BTxB) by mass spectrometry (MS), types C, D, E, and F (BTxC, BTxD, BTxE, BTxF) were now investigated. Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxC, BTxD, BTxE, and BTxF are comprised of a protein complex of the respective neurotoxins with non-toxic non-haemagglutinin (NTNH) and, sometimes, specific haemagglutinins (HA). These protein complexes were observed in mass spectrometric identification. The BTxC complex, from Clostridium botulinum strain 003-9, consisted of a ‘type C1 and D mosaic’ toxin similar to that of type C strain 6813, a non-toxic non-hemagglutinating and a 33kDa hemagglutinating (HA-33) component similar to those of strain C-Stockholm, and an exoenzyme C3 of which the sequence was in full agreement with the known genetic sequence of strain 003-9. The BTxD complex, from C. botulinum strain CB-16, consisted of a neurotoxin with the observed sequence identical with that of type D strain BVD/-3 and of an NTNH with the observed sequence identical with that of type C strain C-Yoichi. Remarkably, the observed protein sequence of CB-16 NTNH differed by one amino acid from the known gene sequence: L859 instead of F859. The BTxE complex, from a C. botulinum isolated from herring sprats, consisted of the neurotoxin with an observed sequence identical with that from strain NCTC 11219 and an NTNH similar to that from type E strain Mashike (1 amino acid difference with observed sequence). BTxF, from C. botulinum strain Langeland (NCTC 10281), consisted of the neurotoxin and an NTNH; observed sequences from both proteins were in agreement with the gene sequence known from strain Langeland. As with BTxA and BTxB, matrix-assisted laser desorption/ionisation (MALDI) MS provided provisional identification from trypsin digest peptide maps and liquid chromatography–electrospray (tandem) mass spectrometry (LC–ES MS) afforded unequivocal identification from amino acid sequence information of digest peptides obtained in trypsin digestion.