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The solubilized full-length KcsA channels with histidine-tagged (His-tag) C-terminal ends were attached to a Ni2+-coated mica surface, and then detergent-destabilized liposomes were added to fill the interchannel space. AFM revealed that the membrane-embedded KcsA channels were oriented with their extracellular faces upward, seen as a tetrameric square shape. This orientation was corroborated by the visible binding of a peptide scorpion toxin, agitoxin-2. To observe the cytoplasmic side of the channel, a His-tag was inserted into the extracellular loop, and the oppositely oriented channels provided wholly different images. In either orientation, the channels were individually dispersed at acidic pH, whereas they were self-assembled at neutral pH, indicating that the oriented channels are allowed to diffuse in the membrane. This method is readily applicable to membrane proteins in general for AFM imaging.<\/jats:p>","DOI":"10.1021\/acs.jpclett.6b03058","type":"journal-article","created":{"date-parts":[[2017,1,31]],"date-time":"2017-01-31T10:13:35Z","timestamp":1485857615000},"page":"785-793","source":"Crossref","is-referenced-by-count":28,"title":["Oriented Reconstitution of the Full-Length KcsA Potassium\nChannel in a Lipid Bilayer for AFM Imaging"],"prefix":"10.1021","volume":"8","author":[{"given":"Ayumi","family":"Sumino","sequence":"first","affiliation":[{"name":"PRESTO, Japan Science and Technology Agency (JST) , 4-1-8 Honcho , , ,","place":["Kawaguchi-shi, Saitama, Japan, 332-0012"]},{"name":"University of Fukui , Faculty of Medical Sciences, , 23-3 Matsuokashimoaizuki , , ,","place":["Yoshida-gun, Fukui, Japan, 910-1193"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takayuki","family":"Uchihashi","sequence":"additional","affiliation":[{"name":"Kanazawa University , , Kakuma-machi, ,","place":["Kanazawa, Japan, 920-1192"]},{"name":"Bio-AFM Frontier Research Center , ,","place":["Kanazawa, Japan, 920-1192"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8438-6750","authenticated-orcid":true,"given":"Shigetoshi","family":"Oiki","sequence":"additional","affiliation":[{"name":"University of Fukui , Faculty of Medical Sciences, , 23-3 Matsuokashimoaizuki , , ,","place":["Yoshida-gun, Fukui, Japan, 910-1193"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2017,1,31]]},"reference":[{"key":"2026090220381825500_cit1","doi-asserted-by":"publisher","first-page":"1451","DOI":"10.1016\/j.bbamem.2013.10.019","article-title":"The Fluid\n- Mosaic Model of Membrane Structure: Still Relevant to Understanding\nthe Structure, Function and Dynamics of Biological Membranes after\nMore than 40 Years","volume":"1838","author":"Nicolson","year":"2014","journal-title":"Biochim. 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