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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

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Figure Lengend Snippet: Effects of valinomycin on K+ accumulation at pH 10. Strains ATCC 9790 (A) and JEM2 (B) were grown in NaTY medium (pH 10), loaded with Na+, and suspended in 0.1 M Na+-CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml. The suspension was (○) or was not (▵) supplemented with 10 mM glucose at 0 min. Valinomycin (30 μM) was added together with glucose at 0 min (●); K+ uptake was initiated by addition of 2 mM KCl at 10 min. The cellular K+ contents were determined by flame photometry.

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

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Figure Lengend Snippet: Time course of changes in DiOC6(3) fluorescence in E. hirae cells. Sodium-loaded cells of ATCC 9790 (A), JEM2 (B), and Nak1 (C) were incubated at a density of 0.5 mg/ml in 0.1 M Na+-CHES (pH 10) with 1 μM DiOC6(3). Fluorescence quenching was initiated by addition of 10 mM glucose (glc) followed by addition of valinomycin (val; 30 μM) and KCl (10 mM). In the experiment shown in panel C, valinomycin and KCl were added simultaneously.

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

Techniques: Fluorescence, Incubation

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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

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Figure Lengend Snippet: K+ accumulation in the presence of valinomycin at various concentrations. Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na+-CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml. The suspension was supplemented with 10 mM glucose and 30 μM valinomycin at −10 min; K+ accumulation was initiated by addition of KCl at various concentrations at 0 min. The established K+ concentration gradients are shown in parentheses. ○, 20 mM KCl; ●, 10 mM KCl; ▵, 5 mM KCl; ▴, 2 mM KCl; □, 1 mM KCl; ■, 0.5 mM KCl; ×, 0.2 mM KCl.

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

Techniques: Cell Culture, Concentration Assay

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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

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Figure Lengend Snippet: Western blotting of cell lysates after denaturing polyacrylamide gel electrophoresis. The cell lysates were prepared from strains ATCC 9790 (lanes 1 and 2) and JEM2 (lanes 3 and 4) grown in NaTY medium (pH 10) as described elsewhere (24). Lysates (5 μg, lanes 1 and 3; 10 μg, lanes 2 and 4) were electrophoresed, immunoblotted with antiserum against purified V1-ATPase (dilution, 1:3,000), and visualized by the alkaline phosphatase system.

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

Techniques: Western Blot, Polyacrylamide Gel Electrophoresis, Purification

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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

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Figure Lengend Snippet: Movements of Na+ and K+ at pH 10. (A and B) Passive Na+ uptake. Sodium uptake was initiated by suspending potassium-loaded cells of strain ATCC 9790 (A) or JEM2 (B) into 200 mM Na+-CHES (pH 10) (●), 180 mM N-methylglucamine-CHES containing 20 mM Na+-CHES (pH 10) (▴), or 200 mM N-methylglucamine-CHES containing 2 mM Na+-CHES (pH 10) (■), respectively. (C and D) ΔΨ-driven Na+ uptake. Potassium-loaded cells of strain ATCC 9790 (C) or JEM2 (D) were suspended in 50 mM N-methylglucamine-CHES (pH 10) buffer containing 2 mM NaCl at −10 min, and the cellular Na+ (triangles) and K+ (circles) contents were monitored; 30 μM valinomycin was added at 0 min (closed symbols).

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

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Article Title: Evidence for Na + Influx via the NtpJ Protein of the KtrII K + Uptake System in Enterococcus hirae

doi:

Figure Lengend Snippet: Effects of valinomycin on the growth of E. hirae at pH 10. Strains ATCC 9790 (A) and JEM2 (B) were cultured in mNaTY (K+-limited NaTY) complex medium at pH 8 (○). At an optical density at 540 nm of 0.08, the medium pH was shifted to 10 by addition of 80 mM Na2CO3 (●). Valinomycin (30 μM) (▵) or KCl (20 mM) (▴) was added 2 min after the addition of the Na2CO3. The cell growth was monitored by cell density measurement.

Article Snippet: Sodium-loaded cells of strains ATCC 9790 (A) and JEM2 (B) cultured in NaTY medium (pH 10) were suspended in 0.1 M Na + -CHES buffer (pH 10) at a cell density equivalent to 1 mg of protein/ml.

Techniques: Cell Culture