Application of protoplast transformation and single- and double-crossover mutagenesis protocols to alkaliphilic Bacillus firmus OF4811M (an auxotrophic strain of B. firmus OF4) facilitated the extension of the sequence of the previously cloned nhaC gene, which encodes an Na+/H+ antiporter, and the surrounding region. The nhaC gene is part of a likely 2-gene operon encompassing nhaC and a small ...

Dogs with compensated biventricular hypertrophy due to chronic atrioventricular block (cAVB), are more susceptible to develop drug-induced Torsade-de-Pointes arrhythmias and sudden cardiac death. It has been suggested that the increased Na(+) influx in hypertrophied cAVB ventricular myocytes contribute to these lethal arrhythmias. The increased Na(+) influx was not mediated by Na(+) channels, i...

Background: NhaA of Escherichia coli can serve as an example for pH regulation and transport mechanism in Na + /H + -exchangers. Results: Different mutations alter Na + and/or H + binding or conformational dynamics. Conclusions: Mutations in different segments of the protein change its pH profile by affecting different aspects of its transport mechanism. Significance: Single site mutations and ...

Na(+)/H(+)-exchangers (NHE) mediate acid extrusion from duodenal epithelial cells, but the isoforms involved have not previously been determined. Thus we investigated 1) the contribution of Na(+)-dependent processes to acid extrusion, 2) sensitivity to Na(+)/H(+) exchange inhibitors, and 3) molecular expression of NHE isoforms. By fluorescence spectroscopy the recovery of intracellular pH (pH(i...

The effect of microenvironmental factors on the regulation of intracellular pH (pHi) in MGH U1 cells and EMT-6 cells was studied using the fluorescent pH probe 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein. Na+/H+ exchange and Na(+)-dependent Cl-/HCO3- exchange were found to be present in both cell types. The activity of both exchangers was dependent on pHi, with low levels of activit...

Functional and immunohistochemical studies were performed to localize and identify Na(+)/H(+) exchanger (NHE) isoforms in macula densa cells. By using the isolated perfused thick ascending limb with attached glomerulus preparation dissected from rabbit kidney, intracellular pH (pH(i)) was measured with fluorescence microscopy by using 2',7'-bis-(2-carboxyethyl)-5-(and -6) carboxyfluorescein. NH...

Most mammalian cells maintain a physiological cytoplasmic pH of approximately 7.2 despite the prediction of a more acidic internal pH on the basis of thermodynamic and metabolic considerations. If the transmembrane gradient of hydrogen ions were determined solely by a hydrogen ion leak pathway in the presence of a membrane potential of -60 mV, then the cytoplasmic pH would be about 6.2, that is...

Fungi possess efficient mechanisms of pH and ion homeostasis, allowing them to grow over a wide range of environmental conditions. In this study, we addressed the role of the pH response transcription factor PacC in salt tolerance of the vascular wilt pathogen Fusarium oxysporum. Loss-of-function pacC(+/-) mutants showed increased sensitivity to Li(+) and Na(+) and accumulated higher levels of ...

A detailed comparison of intracellular pH (pHi) regulatory mechanisms was made between fresh (FBCE) and cultured (CBCE) bovine corneal endothelium to: (1) identify the ion transport mechanisms that could directly or indirectly affect transendothelial HCO3~ transport; and (2) determine if cultured cells could serve as a model for studying transendothelial bicarbonate transport. We used the pH-se...