Cloning of the C-terminal cytoplasmic fragment of the tar protein and effects of the fragment on chemotaxis of Escherichia coli

A gene encoding only the C-terminal portion of the receptor-transducer protein Tar of Escherichia coli was constructed. The gene product was detected and localized in the cytoplasmic fraction of the cell by immunoblotting with anti-Tar antibodies. The C-terminal fragments from wild-type and mutant tar genes were characterized in vivo. The C-terminal fragment generated from tar-526, a mutation that results in a dominant "tumble" phenotype, was found to be deamidated and methylated by the CheB and CheR proteins, respectively. The C-terminal fragment derived from a wild-type gene was poorly deamidated, and the C-terminal fragment derived from tar-529, a dominant mutant with a "smooth swimming" phenotype, was not apparently modified. Cells carrying the C-terminal fragment with the tar-526 mutation as the sole receptor-transducer protein showed a high frequency of tumbling and chemotaxis responses to changes in intracellular pH. These results suggest that the cytoplasmic C-terminal fragment of Tar retains some of the functions of the whole protein in vivo

Effect of Intracellular pH on Rotational Speed of Bacterial Flagellar Motors

Weak acids such as acetate and benzoate, which partially collapse the transmembrane proton gradient, not only mediate pH taxis but also impair the motility of Escherichia coli and Salmonella at an external pH of 5.5. In this study, we examined in more detail the effect of weak acids on motility at various external pH values. A change of external pH over the range 5.0 to 7.8 hardly affected the swimming speed of E. coli cells in the absence of 34 mM potassium acetate. In contrast, the cells decreased their swimming speed significantly as external pH was shifted from pH 7.0 to 5.0 in the presence of 34 mM acetate. The total proton motive force of E. coli cells was not changed greatly by the presence of acetate. We measured the rotational rate of tethered E. coli cells as a function of external pH. Rotational speed decreased rapidly as the external pH was decreased, and at pH 5.0, the motor stopped completely. When the external pH was returned to 7.0, the motor restarted rotating at almost its original level, indicating that high intracellular proton (H(+)) concentration does not irreversibly abolish flagellar motor function. Both the swimming speeds and rotation rates of tethered cells of Salmonella also decreased considerably when the external pH was shifted from pH 7.0 to 5.5 in the presence of 20 mM benzoate. We propose that the increase in the intracellular proton concentration interferes with the release of protons from the torque-generating units, resulting in slowing or stopping of the motors

Nanopatterns induced by pulsed laser irradiation on the surface of an Fe-Al alloy and their magnetic properties

We have studied nanopatterns induced by nanosecond pulsed laser irradiation on (111) plane surfaces of a polycrystalline iron-aluminum alloy and evaluated their magnetic properties. Multiple nanosecond pulsed laser irradiation induces a wavelength-dependent surface transformation of the lattice structure from a B2-type to a supersaturated body centered cubic lattice. The selective formation of surface nanopatterns consisting of holes, stripes, polygonal networks, and dot-like nanoprotrusions can be observed. Furthermore, focused magneto-optical Kerr effect measurements reveal that the magnetic properties of the resultant nanostructured region changes from a paramagnetic to a ferromagnetic phase in accordance with the number of laser pulses

Inhibitions of Histamine Release and Prostaglandin E_2 Synthesis by Mangosteen, a Thai Medicinal Plant

科研費報告書収録論文(課題番号:14370737・基盤研究(B)(2) ・H14～H15/研究代表者:中畑, 則道/Gタンパク質共役型受容体刺激応答の多様性を制御する分子メカニズムに関する研究