POSTERS - BLAST X - University of Utah
POSTERS - BLAST X - University of Utah
POSTERS - BLAST X - University of Utah
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<strong>BLAST</strong> X Poster #25<br />
ATTEMPT TO PURIFY THE HOOK-BASAL BODY WITH C-RING FROM THE Na + -DRIVEN<br />
FLAGELLAR MOTOR<br />
Masafumi Koike, Seiji Kojima, Michio Homma<br />
Division <strong>of</strong> Biological Science, Graduate School <strong>of</strong> Science, Nagoya <strong>University</strong>, Chikusa-ku,<br />
Nagoya 464-8602, Japan.<br />
The flagellum is the biggest architecture in the bacterial organ. At the base <strong>of</strong> each<br />
flagellum, there is a rotary motor embedded in the membrane whose energy source is the<br />
electrochemical gradient <strong>of</strong> the specific ion across the membrane. E. coli and Salmonella have<br />
the H + driven motor, and Vibrio alginolyticus has Na + driven type. The motor is composed <strong>of</strong> the<br />
rotor and the stator, and torque is generated by the interactions between them. The hook-basal<br />
body (HBB) is the rotor part <strong>of</strong> the motor composed <strong>of</strong> rod, LP-, MS-ring and hook. C-ring is<br />
mounted on the cytoplasmic side <strong>of</strong> the MS-ring <strong>of</strong> HBB, and believed to be involved in torque<br />
generation in rotor side <strong>of</strong> the motor. It is composed <strong>of</strong> three proteins; FliM, FliN, and FliG. FliG<br />
is the most closely participated in the torque generation among them. To elucidate the<br />
mechanism <strong>of</strong> torque generation, we undertake the isolation <strong>of</strong> C-ring from the Na + -driven polar<br />
flagellum <strong>of</strong> V. alginolyticus to investigate rotor-stator interactions. We applied the C-ring<br />
isolation method established for Salmonella, but that resulted in the HBB without C-ring. It<br />
indicates that solubilization by Triton X-100 was harsh for C-ring <strong>of</strong> Vibrio. Suitable detergents<br />
were screened and we found that CHAPS could solubilize HBB with FliG attached. Currently<br />
immunoelectron microscopic observation is ongoing to directly detect FliG attached on the MSring.<br />
Also, we are searching more favorable conditions to isolate remaining C-ring proteins, FliM<br />
and FliN.<br />
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