In <i>Escherichia coli</i> Ammonia Inhibits Cytochrome <i>bo</i>₃ But Activates Cytochrome <i>bd</i>-I

• Main Authors: Elena Forte (Author), Sergey A. Siletsky (Author), Vitaliy B. Borisov (Author)
• Format: Book
• Published: MDPI AG, 2020-12-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.
• Notes: 10.3390/antiox10010013; 2076-3921
• Keywords: bacteria; redox enzymes; respiratory oxidases; ammonia; environmental stressor; Therapeutics. Pharmacology; RM1-950
• Source: Antioxidants, Vol 10, Iss 1, p 13 (2020)

Interaction of two redox enzymes of <i>Escherichia coli</i>, cytochrome <i>bo</i>₃ and cytochrome <i>bd</i>-I, with ammonium sulfate/ammonia at pH 7.0 and 8.3 was studied using high-resolution respirometry and absorption spectroscopy. At pH 7.0, the oxygen reductase activity of none of the enzymes is affected by the ligand. At pH 8.3, cytochrome <i>bo</i>₃ is inhibited by the ligand, with 40% maximum inhibition at 100 mM (NH₄)₂SO₄. In contrast, the activity of cytochrome <i>bd</i>-I at pH 8.3 increases with increasing the ligand concentration, the largest increase (140%) is observed at 100 mM (NH₄)₂SO₄. In both cases, the effector molecule is apparently not NH₄⁺ but NH₃. The ligand induces changes in absorption spectra of both oxidized cytochromes at pH 8.3. The magnitude of these changes increases as ammonia concentration is increased, yielding apparent dissociation constants <i>K</i>_d<i>app</i> of 24.3 ± 2.7 mM (NH₄)₂SO₄ (4.9 ± 0.5 mM NH₃) for the Soret region in cytochrome <i>bo</i>₃, and 35.9 ± 7.1 and 24.6 ± 12.4 mM (NH₄)₂SO₄ (7.2 ± 1.4 and 4.9 ± 2.5 mM NH₃) for the Soret and visible regions, respectively, in cytochrome <i>bd</i>-I. Consistently, addition of (NH₄)₂SO₄ to cells of the <i>E. coli</i> mutant containing cytochrome <i>bd</i>-I as the only terminal oxidase at pH 8.3 accelerates the O₂ consumption rate, the highest one (140%) being at 27 mM (NH₄)₂SO₄. We discuss possible molecular mechanisms and physiological significance of modulation of the enzymatic activities by ammonia present at high concentration in the intestines, a niche occupied by <i>E. coli</i>.