Electron cryomicroscopic visualization of PomA/B stator units of the sodium-driven flagellar motor in liposomes.
A motor protein complex of the bacterial flagellum, PomA/B from Vibrio alginolyticus, was reconstituted into liposomes and visualized by electron cryomicroscopy. PomA/B is a sodium channel, composed of two membrane proteins, PomA and PomB, and converts ion flux to the rotation of the flagellar motor. Escherichia coli and Salmonella have a homolog called MotA/B, which utilizes proton instead of sodium ion. PomB and MotB have a peptidoglycan-binding motif in their C-terminal region, and therefore PomA/B and MotA/B are regarded as the stator. Energy filtering electron cryomicroscopy enhanced the image contrast of the proteins reconstituted into liposomes and showed that two extramembrane domains with clearly different sizes stick out of the lipid bilayers on opposite sides. Image analysis combined with gold labeling and deletion of the peptidoglycan-binding motif revealed that the longer one, approximately 70 A long, is likely to correspond to the periplasmic domain, and the other, about half size, to the cytoplasmic domain.
Yonekura
Koji
K
Graduate School of Frontier Biosciences, Osaka University, 1-3, Yamadaoka, Suita, Osaka 565-0871, Japan. yone@msg.ucsf.edu
Yakushi
Toshiharu
T
Atsumi
Tatsuo
T
Maki-Yonekura
Saori
S
Homma
Michio
M
Namba
Keiichi
K
D001426:Bacterial Proteins / Q000378:metabolism / Q000648:ultrastructure*; D020285:Cryoelectron Microscopy; D005407:Flagella / Q000378:metabolism / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008051:Lipid Bilayers; D008081:Liposomes; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000378:metabolism / Q000648:ultrastructure*; D010457:Peptidoglycan / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000378:metabolism / Q000648:ultrastructure*
D001426:Bacterial Proteins; D008051:Lipid Bilayers; D008081:Liposomes; D020409:Molecular Motor Proteins; C063010:MotA protein, Bacteria; C064779:MotB protein, Bacteria; D010457:Peptidoglycan; C107540:PomA protein, Vibrio; C107541:PomB protein, Vibrio; D015222:Sodium Channels; D012964:Sodium