The role of WNK in modulation of KCl cotransport activity in red cells from normal individuals and patients with sickle cell anaemia
| Autor: | John S. Gibson, Anke Hannemann, Rasiqh Wadud, David C. Rees, David C.-Y. Lu, John N. Brewin, Philip S. Low |
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| Přispěvatelé: | Gibson, John S. [0000-0001-6145-9139], Apollo - University of Cambridge Repository, Gibson, John S [0000-0001-6145-9139] |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Erythrocytes Pyrrolidines KCl cotransport Physiology Sodium-Potassium-Chloride Symporters Clinical Biochemistry Anemia Sickle Cell Protein Serine-Threonine Kinases WNK SPAK/OSR1 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Physiology (medical) medicine Staurosporine Humans Protein phosphorylation Sickle cells Phosphorylation Receptor Signaling and cell physiology Red Cell Kinase Chemistry Imidazoles Oxygen tension Cell biology 030104 developmental biology 030220 oncology & carcinogenesis Signal transduction medicine.drug Calyculin Signal Transduction |
| Zdroj: | Pflugers Archiv |
| DOI: | 10.17863/cam.60005 |
| Popis: | Abnormal activity of red cell KCl cotransport (KCC) is involved in pathogenesis of sickle cell anaemia (SCA). KCC-mediated solute loss causes shrinkage, concentrates HbS, and promotes HbS polymerisation. Red cell KCC also responds to various stimuli including pH, volume, urea, and oxygen tension, and regulation involves protein phosphorylation. The main aim of this study was to investigate the role of the WNK/SPAK/OSR1 pathway in sickle cells. The pan WNK inhibitor WNK463 stimulated KCC with an EC50 of 10.9 ± 1.1 nM and 7.9 ± 1.2 nM in sickle and normal red cells, respectively. SPAK/OSR1 inhibitors had little effect. The action of WNK463 was not additive with other kinase inhibitors (staurosporine and N-ethylmaleimide). Its effects were largely abrogated by pre-treatment with the phosphatase inhibitor calyculin A. WNK463 also reduced the effects of physiological KCC stimuli (pH, volume, urea) and abolished any response of KCC to changes in oxygen tension. Finally, although protein kinases have been implicated in regulation of phosphatidylserine exposure, WNK463 had no effect. Findings indicate a predominant role for WNKs in control of KCC in sickle cells but an apparent absence of downstream involvement of SPAK/OSR1. A more complete understanding of the mechanisms will inform pathogenesis whilst manipulation of WNK activity represents a potential therapeutic approach. |
| Databáze: | OpenAIRE |
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