Macrocyclic Peptide-Conjugated Tip for Fast and Selective Molecular Recognition Imaging by High-Speed Atomic Force Microscopy
Macrocyclic Peptide-Conjugated Tip for Fast and Selective Molecular Recognition Imaging by High-Speed Atomic Force Microscopy
复制标题
用于通过高速原子力显微镜进行快速选择性分子识别成像的大环肽缀合尖端
DOI：
10.1021/acsami.1c17708
复制
发表时间：
2021
影响因子：
9.5
通讯作者：
M. Shibata
中科院分区：
文献类型：
--
作者：
L. Pupplin;D. Kanayama;N. Terasaka;K. Sakai;N. Kodera;K. Umeda;A. Sumino;M. Arin;W. Wei;H. Tanaka;T. Fukuma;H. Suga;K. Matsumoto;M. Shibata
在纳米尺度上快速和选择性地识别分子而无需标记是非常期望的，但仍然具有挑战性的目标。在这里，我们展示了使用高速原子力显微镜（HS-AFM）的实时和实时空间识别的未标记的膜受体使用小的合成大环肽共轭的提示。通过对人肝细胞生长因子受体（hMET）的实验验证了单分子识别方法，hMET选择性结合大环肽aMD 4。通过测试和比较用不同长度和刚度的连接体合成的aMD 4，我们最大化了添加到云母表面和支撑的脂质双层的官能化尖端和hMET之间的相互作用。通过HS-AFM的相衬成像使我们能够区分未标记的hMET与不与aMD 4结合的鼠MET同源物。此外，使用小尺寸的配体和连接体，我们实现了同时地形成像的空间分辨率的最小恶化。大环肽在检测无限类型的膜受体具有高选择性和快速成像HS-AFM的多功能性拓宽了未来的应用范围，这种方法的分子识别没有标记。
Fast and selective recognition of molecules at the nanometer scale without labeling is a much desired but still challenging goal to achieve. Here, we show the use of high-speed atomic force microscopy (HS-AFM) for real-time and real-space recognition of unlabeled membrane receptors using tips conjugated with small synthetic macrocyclic peptides. The single-molecule recognition method is validated by experiments on the human hepatocyte growth factor receptor (hMET), which selectively binds to the macrocyclic peptide aMD4. By testing and comparing aMD4 synthesized with linkers of different lengths and rigidities, we maximize the interaction between the functionalized tip and hMET added to both a mica surface and supported lipid bilayers. Phase contrast imaging by HS-AFM enables us to discriminate nonlabeled hMET against the murine MET homologue, which does not bind to aMD4. Moreover, using ligands and linkers of small size, we achieve minimal deterioration of the spatial resolution in simultaneous topographic imaging. The versatility of macrocyclic peptides in detecting unlimited types of membrane receptors with high selectivity and the fast imaging by HS-AFM broaden the range of future applications of this method for molecular recognition without labeling.