title: Impaired Ca2+-induced tyrosine phosphorylation and defective lipid scrambling in erythrocytes from a patient with Scott syndrome: a study using an inhibitor for scramblase that mimics the defect in Scott syndrome.
abstract: Scott syndrome is an hereditary bleeding disorder characterized by a deficiency in platelet procoagulant activity. Unlike normal blood cells, Scott platelets, as well as erythrocytes and lymphocytes, are strongly impaired in their ability to scramble their membrane phospholipids when challenged with Ca2+. In normal cells this collapse of membrane asymmetry leads to surface exposure of phosphatidylserine. Here we report that Scott erythrocytes show an apparent defect in tyrosine phosphorylation on treatment with Ca2+-ionophore. Diminished tyrosine phosphorylation was also apparent in activated Scott platelets, but much less pronounced than observed in red blood cells. On the other hand, tyrosine phosphorylation profiles observed in Scott red blood cell ghosts after sealing in the presence of adenosine triphosphate (ATP) were indistinguishable from those obtained from normal ghosts. Several observations argue in favor of a mechanism in which tyrosine phosphorylation in red blood cells is facilitated by, rather than required for scrambling of membrane lipids. Staurosporin blocks tyrosine phosphorylation in normal red blood cells, but does not inhibit the lipid scrambling process. White ghosts from normal erythrocytes, resealed in the absence of ATP, exhibit Ca2+-induced lipid scrambling without tyrosine phosphorylation. A selective inhibitor of Ca2+-induced lipid scrambling also showed an apparent inhibition of tyrosine phosphorylation in ionophore-treated normal red blood cells, similar to that observed in Scott erythrocytes. While this inhibitor also suppressed Ca2+-induced lipid scrambling in ghosts that were sealed in the presence of ATP, it did not inhibit tyrosine kinase activity. We conclude that the apparent deficiency in tyrosine phosphorylation in Scott cells is an epiphenomenon, possibly associated with a defect in phospholipid scrambling, but not causal to this defect.
authors.affiliation: Department of Biochemistry, Cardiovascular Research Institute Maastricht, Maastricht University, Maastricht, The Netherlands.
authors.forename: D W
authors.initials: DW
authors.lastname: Dekkers
authors.forename: P
authors.initials: P
authors.lastname: Comfurius
authors.forename: W M
authors.initials: WM
authors.lastname: Vuist
authors.forename: J T
authors.initials: JT
authors.lastname: Billheimer
authors.forename: I
authors.initials: I
authors.lastname: Dicker
authors.forename: H J
authors.initials: HJ
authors.lastname: Weiss
authors.forename: R F
authors.initials: RF
authors.lastname: Zwaal
authors.forename: E M
authors.initials: EM
authors.lastname: Bevers
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology; D001778:Blood Coagulation Disorders / Q000097:blood / Q000235:genetics*; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors* / Q000097:blood / Q000235:genetics; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007476:Ionophores / Q000494:pharmacology*; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000097:blood / Q000235:genetics; D008724:Methomyl / Q000031:analogs & derivatives* / Q000494:pharmacology; D010718:Phosphatidylserines / Q000097:blood / Q000502:physiology; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011505:Protein-Tyrosine Kinases / Q000097:blood*; D019311:Staurosporine / Q000494:pharmacology; D013577:Syndrome; D013925:Thromboplastin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D007476:Ionophores; D008563:Membrane Lipids; D008565:Membrane Proteins; D010718:Phosphatidylserines; D049294:Phospholipid Transfer Proteins; D010743:Phospholipids; C111182:ethanimidothioic acid N-((N-butylthio-N-methylamino)-carbonyloxy)-methylester; D008724:Methomyl; D000255:Adenosine Triphosphate; D013925:Thromboplastin; D011505:Protein-Tyrosine Kinases; D019311:Staurosporine; D002118:Calcium