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H+/ATP stoichiometry of cowpea Rhizobium sp. strain 32H1 cells grown under nitrogen-fixing and nitrogen-nonfixing conditions.

The obligate aerobe Cowpea Rhizobium sp. strain 32H1 in axenic culture is able to fix N2 when grown under 0.2% O2 but not when grown under 21% O2. It was, therefore, of interest to investigate ATP synthesis in these cells grown under the two conditions. When respiring in buffers having pHs ranging from 6 to 8.5, cells grown under either O2 tension maintained an intracellular pH more alkaline than the exterior. The transmembrane chemical gradient of H+ (delta pH) was essentially the same under both conditions of growth, decreasing from ca. 90 mV at medium pH 6 to ca. 30 mV at pH 8.5. However, the transmembrane electrical gradient (delta psi) was significantly higher in cells grown under 21% O2 (150 to 166 mV) than in cells grown under 0.2% O2, the latter being 16 mV at pH 6 and increasing to 88 mV at pH 8.5. Therefore, the proton motive force of 21% O2-grown cells ranged from 237 mV at external pH 6 to 185 mV at pH 8.5, compared with a proton motive force of 114 to 121 mV in the 0.2% O2-grown cells. The cells grown in 0.2% O2 had the same proton motive force whether tested at 21 or at 0.2% O2. The phosphorylation potential, calculated from the intracellular ATP, ADP, and Pi concentrations, was 424 mV in the 21% O2-grown cells and 436 mV in the 0.2% O2-grown cells. Thus, the 21% O2-grown cells translocated 1.8 to 2.3 H+/ATP synthesized by the H+-ATPase, whereas the H+/ATP ratio for 0.2% O2-grown cells was 3.7 to 3.8.

Adenosine Triphosphate↗

Reverse nitrogen gradients in the study of phase III and cardiogenic oscillations of the single-breath nitrogen test.

The slope of phase III, phase IV, the slope of phase IV, and cardiac oscillations were measured on tracings obtained by both the regular single-breath N2 test (Tech I) and by a reverse technique (Tech II) in 9 healthy volunteers. Tech II consisted of 3 consecutive vital capacities (VC) of 100% O2 followed by one VC of room air. Theoretically, this should create a reversed apicobasal N2 gradient quantitatively similar to that of Tech I. From the total lung capacity following the VC2 of air, we monitored N2 concentration continuously at the mouth during a slow expiration in a manner similar to that of the single-breath N2 test. With Tech II, it is possible to preserve phase IV and its reversed slope in the presence of an almost flat slope of phase III and markedly blunted cardiac oscillations. When compared to Tech I, the slope of phase III with Tech II decreased from 0.66+/-0.20% N2/L (mean +/-SD) to 0.19+/-0.12 (p is less than 0.001), and cardiac oscillations decreased from a mean % N2 change with each heart beat of 0.87+/-0.37 to 0.24+/-0.21 (p is less than 0.005), whereas phase IV, although reversed in direction, remained quantitatively unchanged (0.35+/-0.15 L with Tech I and 0.37+/-0.14 L with Tech II), and the slope of phase IV tended to increase (2.7+/-1.9% N2 with Tech I and 3.4+/-2.1% N2 with Tech II, p=NS). We conclude that the N2 gradients within the lungs responsible for the slope of phase III and cardiac oscillations are largely independent of the gradients that give rise to phase IV and the slope of phase IV.

Adult↗