From the mouse to the mass spectrometer: Detection and differentiation of the endoproteinase activities of botulinum neurotoxins A-G by mass spectrometry
From the mouse to the mass spectrometer: Detection and differentiation of the endoproteinase activities of botulinum neurotoxins A-G by mass spectrometry
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DOI：
10.1021/ac050485f
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发表时间：
2005-07-01
影响因子：
7.4
通讯作者：
Barr, JR
中科院分区：
文献类型：
--
作者：
Boyer, AE;Moura, H;Barr, JR
我们已经开发了一种测定（Endopep-MS），通过质谱（MS）检测所有七种BoNT类型的特异性内切蛋白酶活性。每种BoNT类型都切割参与神经元传递的蛋白质上的独特位点。基于这些蛋白质的靶肽底物通过其对底物的酶促作用和两种肽产物的产生来识别BoNT类型，然后通过基质辅助激光解吸/电离飞行时间MS或液相色谱电喷雾电离MS/MS来检测。我们显示了在多重检测格式中检测所有七种毒素类型的能力。在缓冲系统中，A、B、E和F型毒素的检测限范围为0.039至0.625小鼠LD 50/mL。Endopep-MS分析是第一个区分所有七种BoNT类型的方法，具有灵敏度，特异性，并有可能定量毒素活性。
We have developed an assay (Endopep-MS) that detects the specific endoproteinase activities of all seven BoNT types by mass spectrometry (MS). Each BoNT type cleaves a unique site on proteins involved in neuronal transmission. Target peptide substrates based on these proteins identify a BoNT type by its enzymatic action on the substrate and the production of two peptide products, which are then detected by matrix-assisted laser desorption/ionization time-of-flight MS or liquid chromatography electrospray ionization MS/MS. We showed the ability to detect all seven toxin types in a multiplexed assay format. The detection limits achieved range from 0.039 to 0.625 mouse LD50/mL for toxin types A, B, E, and F in a buffer system. The Endopep-MS assay is the first to differentiate all seven BoNT types, is sensitive, specific, and has the potential to quantify toxin activity.