TY - JOUR
T1 - Chained Structure of Dimeric F1-like ATPase in Mycoplasma mobile Gliding Machinery
AU - Toyonaga, Takuma
AU - Kato, Takayuki
AU - Kawamoto, Akihiro
AU - Kodera, Noriyuki
AU - Hamaguchi, Tasuku
AU - Tahara, Yuhei O.
AU - Ando, Toshio
AU - Namba, Keiichi
AU - Miyata, Makoto
N1 - Funding Information:
We thank Toshiaki Arata, Ikuko Fujiwara, Kohei Kobayashi, and Hiroki Sato at the Graduate School of Science, Osaka City University, and Takayuki Uchihashi at the Department of Physics and Structural Biology Research Center, Nagoya University, for helpful discussions. We also thank Aya Takamori at the Graduate School of Science, Osaka City University, for performing matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry. T.T. learned RELION software in an instruction course on 20180927 to 28 provided by the Cyclic Innovation for Clinical Empowerment (CiCLE) from AMED. This work was supported by a grant-in-aid for scientific research on the Innovative Area Harmonized Supramolecular Motility Machinery and Its Diversity (MEXT KAKENHI grant number JP24117002), by grants-in-aid for scientific research (B) and (A) (MEXT KAKENHI grant numbers JP24390107 and JP17H01544), by JST CREST grant number JPMJCR19S5, Japan, by the Osaka City University (OCU) Strategic Research Grant 2018 for top priority researches, and by a grant-in-aid of the Fugaku Trust for Medicinal Research to M.M. and JSPS KAKENHI (grant number JP25000013), the Platform Project for Supporting Drug Discovery and Life Science Research (BINDS) from AMED (grant number JP19am0101117 and support number 1282), CiCLE (grant number JP17pc0101020), and JEOL YOKOGUSHI Research Alliance Laboratories of Osaka University to K.N.
Funding Information:
This work was supported by a grant-in-aid for scientific research on the Innovative Area Harmonized Supramolecular Motility Machinery and Its Diversity (MEXT KAKENHI grant number JP24117002), by grants-in-aid for scientific research (B) and (A) (MEXT KAKENHI grant numbers JP24390107 and JP17H01544), by JST CREST grant number JPMJCR19S5, Japan, by the Osaka City University (OCU) Strategic Research Grant 2018 for top priority researches, and by a grant-in-aid of the Fugaku Trust for Medicinal Research to M.M. and JSPS KAKENHI (grant number JP25000013), the Platform Project for Supporting Drug Discovery and Life Science Research (BINDS) from AMED (grant number JP19am0101117 and support number 1282), CiCLE (grant number JP17pc0101020), and JEOL YOKOGUSHI Research Alliance Laboratories of Osaka University to K.N.
Publisher Copyright:
© 2021 Toyonaga et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license
PY - 2021/8/1
Y1 - 2021/8/1
N2 - Mycoplasma mobile, a fish pathogen, exhibits gliding motility using ATP hydrolysis on solid surfaces, including animal cells. The gliding machinery can be divided into surface and internal structures. The internal structure of the motor is composed of 28 so-called “chains” that are each composed of 17 repeating protein units called “particles.” These proteins include homologs of the catalytic a and b subunits of F1-ATPase. In this study, we isolated the particles and determined their structures using negative-staining electron microscopy and high-speed atomic force microscopy. The isolated particles were composed of five proteins, MMOB1660 (a-subunit homolog), -1670 (b-subunit homolog), -1630, -1620, and -4530, and showed ATP hydrolyzing activity. The two-dimensional (2D) structure, with dimensions of 35 and 26nm, showed a dimer of hexameric ring approximately 12nm in diameter, resembling F1-ATPase catalytic (ab)3. We isolated the F1-like ATPase unit, which is composed of MMOB1660, -1670, and -1630. Furthermore, we isolated the chain and analyzed the three-dimensional (3D) structure, showing that dimers of mushroom-like structures resembling F1-ATPase were connected and aligned along the dimer axis at 31-nm intervals. An atomic model of F1-ATPase catalytic (ab)3 from Bacillus PS3 was successfully fitted to each hexameric ring of the mushroom-like structure. These results suggest that the motor for M. mobile gliding shares an evolutionary origin with F1-ATPase. Based on the obtained structure, we propose possible force transmission processes in the gliding mechanism. IMPORTANCE F1Fo-ATPase, a rotary ATPase, is widespread in the membranes of mitochondria, chloroplasts, and bacteria and converts ATP energy with a proton motive force across the membrane by its physical rotation. Homologous protein complexes play roles in ion and protein transport. Mycoplasma mobile, a pathogenic bacterium, was recently suggested to have a special motility system evolutionarily derived from F1-ATPase. The present study isolated the protein complex from Mycoplasma cells and supported this conclusion by clarifying the detailed structures containing common and novel features as F1-ATPase relatives.
AB - Mycoplasma mobile, a fish pathogen, exhibits gliding motility using ATP hydrolysis on solid surfaces, including animal cells. The gliding machinery can be divided into surface and internal structures. The internal structure of the motor is composed of 28 so-called “chains” that are each composed of 17 repeating protein units called “particles.” These proteins include homologs of the catalytic a and b subunits of F1-ATPase. In this study, we isolated the particles and determined their structures using negative-staining electron microscopy and high-speed atomic force microscopy. The isolated particles were composed of five proteins, MMOB1660 (a-subunit homolog), -1670 (b-subunit homolog), -1630, -1620, and -4530, and showed ATP hydrolyzing activity. The two-dimensional (2D) structure, with dimensions of 35 and 26nm, showed a dimer of hexameric ring approximately 12nm in diameter, resembling F1-ATPase catalytic (ab)3. We isolated the F1-like ATPase unit, which is composed of MMOB1660, -1670, and -1630. Furthermore, we isolated the chain and analyzed the three-dimensional (3D) structure, showing that dimers of mushroom-like structures resembling F1-ATPase were connected and aligned along the dimer axis at 31-nm intervals. An atomic model of F1-ATPase catalytic (ab)3 from Bacillus PS3 was successfully fitted to each hexameric ring of the mushroom-like structure. These results suggest that the motor for M. mobile gliding shares an evolutionary origin with F1-ATPase. Based on the obtained structure, we propose possible force transmission processes in the gliding mechanism. IMPORTANCE F1Fo-ATPase, a rotary ATPase, is widespread in the membranes of mitochondria, chloroplasts, and bacteria and converts ATP energy with a proton motive force across the membrane by its physical rotation. Homologous protein complexes play roles in ion and protein transport. Mycoplasma mobile, a pathogenic bacterium, was recently suggested to have a special motility system evolutionarily derived from F1-ATPase. The present study isolated the protein complex from Mycoplasma cells and supported this conclusion by clarifying the detailed structures containing common and novel features as F1-ATPase relatives.
KW - Atomic force microscopy
KW - Bacterial motility
KW - Electron microscopy
KW - F-ATPase
KW - Parasitic bacteria
KW - Rotary motor
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U2 - 10.1128/mBio.01414-21
DO - 10.1128/mBio.01414-21
M3 - Article
C2 - 34281395
AN - SCOPUS:85115449296
SN - 2161-2129
VL - 12
JO - mBio
JF - mBio
IS - 4
M1 - e01414-21
ER -