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J Biochem. 2013 May;153(5):441-51. doi: 10.1093/jb/mvt011. Epub 2013 Feb 18.

Na+ conductivity of the Na+-driven flagellar motor complex composed of unplugged wild-type or mutant PomB with PomA.

Author information

1
Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan.

Abstract

PomA and PomB form the stator complex, which functions as a Na(+) channel, in the Na(+)-driven flagellar motor of Vibrio alginolyticus. The plug region of PomB is thought to regulate the Na(+) flow and to suppress massive ion influx through the stator channel. In this study, in order to measure the Na(+) conductivity of the unplugged stator, we over-produced a plug-deleted stator of the Na(+)-driven flagellar motor in Escherichia coli. The over-production of the plug-deleted stator in E. coli cells caused more severe growth inhibition than in Vibrio cells and that growth inhibition depended on the Na(+) concentration in the growth medium. Measurement of intracellular Na(+) concentration by flame photometry and fluorescent analysis with a Na(+) indicator, Sodium Green, revealed that over-production of the plug-deleted stator increased the Na(+) concentration in cell. Some mutations in the channel region of PomB or in the cytoplasmic region of PomA suppressed both the growth inhibition and the increase in intracellular Na(+) concentration. These results suggest that the level of growth inhibition correlates with the intracellular Na(+) concentration, probably due to the Na(+) conductivity through the stator due to the mutations.

PMID:
23420849
DOI:
10.1093/jb/mvt011
[Indexed for MEDLINE]

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