Macrocyclic Peptide-Conjugated Tip for Fast and Selective Molecular Recognition Imaging by High-Speed Atomic Force Microscopy
mediaposted on 2021-11-12, 13:34 authored by Leonardo Puppulin, Daiki Kanayama, Naohiro Terasaka, Katsuya Sakai, Noriyuki Kodera, Kenichi Umeda, Ayumi Sumino, Arin Marchesi, Wei Weilin, Hideo Tanaka, Takeshi Fukuma, Hiroaki Suga, Kunio Matsumoto, Mikihiro Shibata
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.
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supported lipid bilayersstill challenging goalmurine met homologuedetecting unlimited typessimultaneous topographic imagingphase contrast imagingdiscriminate nonlabeled hmetcomparing amd4 synthesizedafm enables usachieve minimal deteriorationmolecule recognition methodmacrocyclic peptide amd4macrocyclic peptidespace recognitionselective recognitionmacrocyclic peptideshmet addedhmet ),fast imagingusing ligandsspatial resolutionsmall sizeselectively bindsmuch desiredmica surfacemembrane receptorsfuture applicationsfunctionalized tipdifferent lengthsconjugated tipafm broaden