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Title: Elastic rotation of Escherichia coli F{sub O}F{sub 1} having ε subunit fused with cytochrome b{sub 562} or flavodoxin reductase

Highlights: • Intra-molecular rotation of F{sub O}F{sub 1} ATP synthase was observed using a small bead probe. • Carboxyl-terminus of the ε subunit was fused to cytochrome b{sub 562} or flavodoxin reductase. • The F{sub O}F{sub 1} showed continual rotation with similar rate to the wild-type enzyme. • The intra-molecular rotation is flexible and elastic. - Abstract: Intra-molecular rotation of F{sub O}F{sub 1} ATP synthase enables cooperative synthesis and hydrolysis of ATP. In this study, using a small gold bead probe, we observed fast rotation close to the real rate that would be exhibited without probes. Using this experimental system, we tested the rotation of F{sub O}F{sub 1} with the ε subunit connected to a globular protein [cytochrome b{sub 562} (ε-Cyt) or flavodoxin reductase (ε-FlavR)], which is apparently larger than the space between the central and the peripheral stalks. The enzymes containing ε-Cyt and ε-FlavR showed continual rotations with average rates of 185 and 148 rps, respectively, similar to the wild type (172 rps). However, the enzymes with ε-Cyt or ε-FlavR showed a reduced proton transport. These results indicate that the intra-molecular rotation is elastic but proton transport requires more strict subunit/subunit interaction.
Authors:
 [1] ; ;  [2] ;  [3] ;  [2] ;  [1]
  1. Department of Bioscience, Nagahama Institute of Bioscience and Technology, Nagahama, Shiga 526-0829 (Japan)
  2. Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Yahaba, Iwate 028-3694 (Japan)
  3. Department of Biochemistry, University of Western Ontario, London, Ontario N6A 5C1 (Canada)
Publication Date:
OSTI Identifier:
22416376
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 446; Journal Issue: 4; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ATP; ENZYMES; ESCHERICHIA COLI; ETHERS; GLYCINE; GOLD; HYDROLYSIS; PROBES; PROTON TRANSPORT; ROTATION; SYNTHESIS