Random phage display based screening of peptides that bind to botulinum neurotoxin binding protein, nontoxic nonhemagglutinin.
Random phage display based screening of peptides that bind to botulinum neurotoxin binding protein, nontoxic nonhemagglutinin.
复制标题
基于随机噬菌体展示筛选与肉毒杆菌神经毒素结合蛋白、无毒非血凝素结合的肽。
DOI：
10.1007/s00284-013-0355-0
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发表时间：
2013
影响因子：
2.6
通讯作者：
Niwa K,
中科院分区：
文献类型：
--
作者：
Sagane Y;Miyata K;Kurihara S;Yoneyama T;Inui K;Miyashita S;Hayashi S;Suzuki T;Niwa K,
肉毒神经毒素（BoNT）以pH依赖的方式与无毒的非血凝素（NTNHA）蛋白结合，并产生抗蛋白酶的BoNT/NTNHA复合物。在这里，我们使用随机噬菌体展示技术筛选与血清型D NTNHA（NTNHA-D）结合的短肽。将NTNHA固定在石英晶体微天平（QCM）电极上，在酸性条件下将NTNHA-D暴露于展示随机七肽的底物中。在用酸性缓冲液冲洗后，收集在碱性条件下释放的细菌。通过QCM上的频移监测噬菌体的结合和释放。作为筛选的结果，选择了16种作为与NTNHA-D结合的肽。所选择的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。这将解释BoNT与NTNHA蛋白的结合方式。
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.