Elastic rotation of Escherichia coli F(O)F(1) having ε subunit fused with cytochrome b(562) or flavodoxin reductase

Hideyuki Oka; Hiroyuki Hosokawa; Mayumi Nakanishi-Matsui; Stanley D Dunn; Masamitsu Futai; Atsuko Iwamoto-Kihara

doi:10.1016/j.bbrc.2014.03.021

Elastic rotation of Escherichia coli F(O)F(1) having ε subunit fused with cytochrome b(562) or flavodoxin reductase

Biochem Biophys Res Commun. 2014 Apr 18;446(4):889-93. doi: 10.1016/j.bbrc.2014.03.021. Epub 2014 Mar 13.

Authors

Hideyuki Oka¹, Hiroyuki Hosokawa², Mayumi Nakanishi-Matsui², Stanley D Dunn³, Masamitsu Futai², Atsuko Iwamoto-Kihara⁴

Affiliations

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.
⁴ Department of Bioscience, Nagahama Institute of Bioscience and Technology, Nagahama, Shiga 526-0829, Japan. Electronic address: a_iwamoto@nagahama-i-bio.ac.jp.

PMID: 24631905
DOI: 10.1016/j.bbrc.2014.03.021

Abstract

Intra-molecular rotation of FOF1 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 FOF1 with the ε subunit connected to a globular protein [cytochrome b562 (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.

Keywords: ATP synthase; Elastic rotation; Energy coupling; F(O)F(1); Molecular motor; ε Subunit.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Artificial Gene Fusion
Cytochrome b Group / genetics
Cytochrome b Group / metabolism*
Escherichia coli / chemistry
Escherichia coli / genetics
Escherichia coli / metabolism*
Escherichia coli Proteins / genetics
Escherichia coli Proteins / metabolism*
Gene Fusion*
Models, Molecular
NADH, NADPH Oxidoreductases / genetics
NADH, NADPH Oxidoreductases / metabolism*
Protein Conformation
Protein Subunits / chemistry
Protein Subunits / genetics
Protein Subunits / metabolism
Proton-Translocating ATPases / chemistry
Proton-Translocating ATPases / genetics
Proton-Translocating ATPases / metabolism*

Substances

Cytochrome b Group
Escherichia coli Proteins
Protein Subunits
cytochrome b562, E coli
NADH, NADPH Oxidoreductases
flavodoxin NADPH oxidoreductase
Proton-Translocating ATPases