Reversible control of F1-ATPase rotational motion using a photochromic ATP analog at the single molecule level |
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Authors: | Ei-Ichiro Sunamura Takashi Kamei Hiroki Konno Nobuyuki Tamaoki Toru Hisabori |
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Affiliation: | 1. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Chiyoda-Ku Tokyo 102-0076, Japan;2. Chemical Resources Laboratory, Tokyo Institute of Technology, Nagatsuta 4259-R1-8, Midori-Ku, Yokohama 226-8503, Japan;3. Research Institute for Electronic Science, Hokkaido University, N20, W10, Kita-Ku, Sapporo 001-0020, Japan;4. Imaging Research Division, Bio-AFM Frontier Research Center, Kanazawa University, Kakuma, Kanazawa 920-1192, Japan |
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Abstract: | Motor enzymes such as F1-ATPase and kinesin utilize energy from ATP for their motion. Molecular motions of these enzymes are critical to their catalytic mechanisms and were analyzed thoroughly using a single molecule observation technique. As a tool to analyze and control the ATP-driven motor enzyme motion, we recently synthesized a photoresponsive ATP analog with a p-tert-butylazobenzene tethered to the 2′ position of the ribose ring. Using cis/trans isomerization of the azobenzene moiety, we achieved a successful reversible photochromic control over a kinesin-microtubule system in an in vitro motility assay. Here we succeeded to control the hydrolytic activity and rotation of the rotary motor enzyme, F1-ATPase, using this photosensitive ATP analog. Subsequent single molecule observations indicated a unique pause occurring at the ATP binding angle position in the presence of cis form of the analog. |
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Keywords: | FoF1, FoF1-ATP synthase F1 (F1-ATPase), catalytic moiety of ATP synthase Fo, membrane sector of ATP synthase TNP-ATP, 2&prime ,3&prime -O-(2,4,6-trinitrophenyl) adenosine 5&prime -triphosphate AMP-PNP, adenylyl imidodiphosphate VIS light, visible light ATP-AzotBu, ATP analog containing azobenzene at the 2&prime position of the ribose and a tert-butyl group at the azobenzene terminus trans-ATP-AzotBu, trans form of ATP-AzotBu cis-ATP-AzotBu, cis form of ATP-AzotBu PSS, photostationary state |
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