title: Identification of the Ca²⁺ entry pathway involved in deoxygenation-induced phosphatidylserine exposure in red blood cells from patients with sickle cell disease.
abstract: Phosphatidylserine (PS) exposure in red blood cells (RBCs) from sickle cell disease (SCD) patients is increased compared to levels in normal individuals and may participate in the anaemic and ischaemic complications of SCD. Exposure is increased by deoxygenation and occurs with elevation of intracellular Ca²⁺ to low micromolar levels. The Ca²⁺ entry step has not been defined but a role for the deoxygenation-induced pathway, Psickle, is postulated. Partial Psickle inhibitors 4-acetamido-4'-isothiocyanostilbene-2,2'-disulphonic acid (SITS), 4,4'-dithiocyano-2,2'-stilbene-disulphonic acid (DIDS) and dipyridamole inhibited deoxygenation-induced PS exposure (DIDS IC50, 118 nM). Inhibitors and activators of other pathways (including these stimulated by depolarisation, benzodiazepines, glutamate and stretch) were without effect. Zn²⁺ and Gd³⁺ stimulated PS exposure to high levels. In the case of Zn²⁺, this effect was independent of oxygen (and hence HbS polymerisation and RBC sickling) but required extracellular Ca²⁺. The effect was completely abolished when Zn²⁺ (100 μM) was added to RBCs suspended in autologous plasma, implying a requirement of high levels of free Zn²⁺.
authors.affiliation: Department of Veterinary Medicine, Madingley Road, Cambridge, CB3 0ES, UK.
authors.forename: U M
authors.initials: UM
authors.lastname: Cytlak
authors.forename: A
authors.initials: A
authors.lastname: Hannemann
authors.forename: D C
authors.initials: DC
authors.lastname: Rees
authors.forename: J S
authors.initials: JS
authors.lastname: Gibson
mesh_terms: D000755:Anemia, Sickle Cell / Q000097:blood / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D004913:Erythrocytes, Abnormal / Q000187:drug effects / Q000378:metabolism*; D005682:Gadolinium / Q000494:pharmacology; D006451:Hemoglobin, Sickle / Q000378:metabolism; D006801:Humans; D010100:Oxygen / Q000494:pharmacology*; D010718:Phosphatidylserines / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
chemical_list: D006451:Hemoglobin, Sickle; D010718:Phosphatidylserines; D005682:Gadolinium; D015032:Zinc; D010100:Oxygen; D002118:Calcium