title: Protein kinase CK1α regulates erythrocyte survival.
abstract: Protein kinase CK1 (casein kinase 1) isoforms are involved in the regulation of various physiological functions including apoptosis. The specific CK1 inhibitor D4476 may either inhibit or foster apoptosis. Similar to apoptosis of nucleated cells, eryptosis, the suicidal death of erythrocytes, is paralleled by cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the cell surface. Triggers of eryptosis include increase of cytosolic Ca(2+) activity following energy depletion (removal of glucose) or oxidative stress (exposure to the oxidant tert-butyl hydroperoxide [TBOOH]). Western blotting was utilized to verify that erythrocytes express the protein kinase CK1α, and FACS analysis to determine whether the CK1 inhibitor D4476 and CK1α activator pyrvinium pamoate modify forward scatter (reflecting cell volume), annexin V binding (reflecting phosphatidylserine exposure), and Fluo3 fluorescence (reflecting cytosolic Ca(2+) activity). As a result, both, human and murine erythrocytes express CK1 isoform α. Glucose depletion (48 hours) and exposure to 0.3 mM TBOOH (30 minutes) both decreased forward scatter, increased annexin V binding and increased Fluo3 fluorescence. CK1 inhibitor D4476 (10 μM) significantly blunted the decrease in forward scatter, the increase in annexin V binding and the increase in Fluo 3 fluorescence. (R)-DRF053, another CK1 inhibitor, similarly blunted the increase in annexin V binding upon glucose depletion. The CK1α specific activator pyrvinium pamoate (10 μM) significantly enhanced the increase in annexin V binding and Fluo3 fluorescence upon glucose depletion and TBOOH exposure. In the presence of glucose, pyrvinium pamoate slightly but significantly increased Fluo3 fluorescence. In conclusion, CK1 isoform α participates in the regulation of erythrocyte programmed cell death by modulating cytosolic Ca(2+) activity.
authors.affiliation: Department of Physiology, University of Tübingen, Tübingen, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Zelenak
authors.forename: Matthias
authors.initials: M
authors.lastname: Eberhard
authors.forename: Kashif
authors.initials: K
authors.lastname: Jilani
authors.forename: Syed M
authors.initials: SM
authors.lastname: Qadri
authors.forename: Boris
authors.initials: B
authors.lastname: Macek
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D001549:Benzamides / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D048128:Casein Kinase Ialpha / Q000037:antagonists & inhibitors / Q000378:metabolism*; D048429:Cell Size / Q000187:drug effects; D004912:Erythrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D051379:Mice; D010718:Phosphatidylserines / Q000494:pharmacology; D011774:Pyrvinium Compounds / Q000494:pharmacology; D014966:Xanthenes / Q000737:chemistry; D020122:tert-Butylhydroperoxide / Q000494:pharmacology
chemical_list: C493177:4-(4-(2,3-dihydrobenzo(1,4)dioxin-6-yl)-5-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D000814:Aniline Compounds; D017304:Annexin A5; D001549:Benzamides; D007093:Imidazoles; D010718:Phosphatidylserines; D011774:Pyrvinium Compounds; D014966:Xanthenes; C059715:Fluo-3; C024631:pyrvinium; D020122:tert-Butylhydroperoxide; D048128:Casein Kinase Ialpha; D005947:Glucose; D002118:Calcium