Crystallization and preliminary X-ray analysis of MotY, a stator component of the Vibrio alginolyticus polar flagellar motor.
The polar flagellum of Vibrio alginolyticus is rotated by the sodium motor. The stator unit of the sodium motor consists of four different proteins: PomA, PomB, MotX and MotY. MotX and MotY, which are unique components of the sodium motor, form the T-ring structure attached to the LP ring in the periplasmic space. MotY has a putative peptidoglycan-binding motif in its C-terminal region and MotX is suggested to interact with PomB. Thus, MotX and MotY are thought to be required for incorporation and stabilization of the PomA/B complex. In this study, mature MotY composed of 272 amino-acid residues and its SeMet derivative were expressed with a C-terminal hexahistidine-tag sequence, purified and crystallized. Native crystals were grown in the hexagonal space group P6(1)22/P6(5)22, with unit-cell parameters a = b = 104.1, c = 132.6 A. SeMet-derivative crystals belonged to the same space group with the same unit-cell parameters as the native crystals. Anomalous difference Patterson maps of the SeMet derivative showed significant peaks in their Harker sections, indicating that the derivatives are suitable for structure determination.
Shinohara
Akari
A
Soft Nano-Machine Project, CREST, JST, Chikusa-ku, Nagoya 464-8602, Japan.
Sakuma
Mayuko
M
Yakushi
Toshiharu
T
Kojima
Seiji
S
Namba
Keiichi
K
Homma
Michio
M
Imada
Katsumi
K
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000737:chemistry; D020409:Molecular Motor Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics; D044149:Vibrio alginolyticus / Q000737:chemistry* / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D020409:Molecular Motor Proteins; C088135:MotY protein, Vibrio