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Abstract Nitrite uptake into red blood cells (RBCs) precedes its intracellular reactions with hemoglobin (Hb) that forms nitric oxide (NO) during hypoxia. We investigated the uptake of nitrite and its reactions with Hb at different oxygen saturations (S o 2 ), using RBCs with (carp and rabbit) and without (hagfish and lamprey) anion exchanger-1 (AE1) in the membrane, with the aim to unravel the mechanisms and oxygenation dependencies of nitrite transport. Added nitrite rapidly diffused into the RBCs until equilibrium. The distribution ratio of nitrite across the membrane agreed with that expected from HNO 2 diffusion and AE1-mediated facilitated NO 2 − diffusion. Participation of HNO 2 diffusion was emphasized by rapid transmembrane nitrite equilibration also in the natural AE1 knockouts. Following the equilibration, nitrite was consumed by reacting with Hb, which created a continued inward diffusion controlled by intracellular reaction rates. Changes in nitrite uptake with S o 2 , pH, or species were accordingly explained by corresponding changes in reaction rates. In carp, nitrite uptake rates increased linearly with decreasing S o 2 over the entire S o 2 range. In rabbit, nitrite uptake rates were highest at intermediate S o 2 , producing a bell-shaped relationship with S o 2 . Nitrite consumption increased ∼10-fold with a 1 unit decrease in pH, as expected from the involvement of protons in the reactions with Hb. The reaction of nitrite with deoxyhemoglobin was favored over that with oxyhemoglobin at intermediate S o 2 . We propose a model for RBC nitrite uptake that involves both HNO 2 diffusion and AE1-mediated transport and that explains both the present and previous (sometimes puzzling) results. anion exchanger-1 facilitated diffusion HNO 2 diffusion nitric oxide Copyright © 2010 the American Physiological Society
AJP - Regulatory, Integrative and Comparative Physiology – The American Physiological Society
Published: Apr 1, 2010
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