Abstract
The effects of oxygen tension (PO 2) upon the K influx pathways of equine red cells have been studied using 86Rb+ as congener for K. Equilibration of cells in 100% nitrogen led to a low and Cl-independent K flux. Change to an atmosphere of 100% air led to a rapid sixfold increase in K flux. The oxygen-activated flux was entirely Cl dependent and was maintained for up to 3 h. Oxygenation-evoked activation was dependent upon PO 2 over the physiological range with little effect up to 70% saturation of haemoglobin with oxygen but significant effects between 70 and 100%. K flux at low PO 2 was unaffected by acidification to pH 7 or by hypotonic cell swelling. By contrast, at high PO 2 both manipulations caused a substantial increase in Cl-dependent K flux. N-Ethylmaleimide (NEM; 1 mM) caused a progressive activation of KC1 cotransport in cells held under nitrogen. The protein phosphatase inhibitor, calyculin A (100 nM), applied during NEM-evoked activation caused a “clamping” of K influx at that level. This “clamped” activity was unaffected by subsequent oxygenation. We conclude that oxygenation exerts a primary control over cotransport activity and that acidification and cell swelling are secondary modulators. It appears that oxygenation-evoked activation of the Cl-dependent K flux involves a serine/threonine phosphorylation event. Regulating the PO 2 of the solution before and during experiments is important in controlling the activity of the KC1 cotransporter and cell volume.
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Honess, N.A., Gibson, J.S. & Cossins, A.R. The effects of oxygenation upon the Cl-dependent K flux pathway in equine red cells. Pflugers Arch. 432, 270–277 (1996). https://doi.org/10.1007/s004240050133
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DOI: https://doi.org/10.1007/s004240050133