Department of Food and Cosmetic Science, Faculty of Bioindustry, Tokyo University of Agriculture, 196 Yasaka, Abashiri, 099-2493, Japan.
Curr Microbiol. 2013 Aug;67(2):188-92. doi: 10.1007/s00284-013-0355-0. Epub 2013 Mar 16.
Botulinum neurotoxin (BoNT) binds to nontoxic nonhemagglutinin (NTNHA) protein in a pH-dependent manner, and yields the protease-resistant BoNT/NTNHA complex. Here, we screened short peptides that bind to the serotype D NTNHA (NTNHA-D) using random phage display technique. NTNHA was fixed onto electrode of quartz crystal microbalance (QCM) apparatus, and then the phages displaying random heptapeptides were exposed to the NTNHA-D under the acidic condition. After rinsing with acidic buffer, the released phages under the alkaline condition were collected. The binding and release of the phage were monitored by the frequency shift on the QCM. As a result of the screening, 16 were selected as peptides that bind to NTNHA-D. The selected peptides do not share any conserved sequence, but tend to be rich in basic and/or hydrophobic amino acid. This would explain the binding manner of the BoNT to the NTNHA protein.
肉毒神经毒素（BoNT）以依赖 pH 的方式与非毒性非血凝素（NTNHA）蛋白结合，并产生蛋白酶抗性的 BoNT/NTNHA 复合物。在这里，我们使用随机噬菌体展示技术筛选了与血清型 D NTNHA（NTNHA-D）结合的短肽。将 NTNHA 固定在石英晶体微天平（QCM）仪器的电极上，然后将展示随机七肽的噬菌体在酸性条件下暴露于 NTNHA-D。用酸性缓冲液冲洗后，在碱性条件下收集释放的噬菌体。通过 QCM 上的频率偏移监测噬菌体的结合和解离。经过筛选，选择了 16 个与 NTNHA-D 结合的肽。所选肽不共享任何保守序列，但倾向于富含碱性和/或疏水性氨基酸。这可以解释 BoNT 与 NTNHA 蛋白的结合方式。
