Subject(s)
Amino Acid Transport Systems, Neutral/genetics , Blood Group Antigens/blood , Blood Group Antigens/genetics , Erythrocytes/metabolism , Hemoglobinuria, Paroxysmal/blood , Hemoglobinuria, Paroxysmal/genetics , Adult , Aged , Amino Acid Transport Systems, Neutral/blood , Female , Flow Cytometry/methods , Hemoglobinuria, Paroxysmal/diagnosis , Humans , Male , Middle Aged , Young AdultABSTRACT
XK is a putative transporter of unknown function that is ubiquitously expressed and linked through disulfide bonds to Kell protein, an endothelin-3 (ET-3)-converting enzyme. We generated three knockout (KO) mice that lacked either Xk, Kell or both proteins and characterized erythrocyte cation levels, transport and hematological parameters. Absence of Xk or Kell was accompanied by changes in erythrocyte K(+), Mg(2+), Na(+) and Ca(2+) transport that were associated with changes in mean cellular volume and corpuscular hemoglobin concentration mean. Baseline Ca(2+)-ATPase activity was undetected in erythrocytes from all three mouse types but was restored upon pre-incubation with ET-3. Consistent with these alterations in Ca(2+) handling, we observed increased Gardos channel activity in Kel and Xk KO mice. In addition Kel deletion was associated with increased Mg(2+) permeability while Xk deletion blocked Na/Mg exchanger activity. Our results provide evidence that cellular divalent cation regulation is functionally coupled to the Kell/XK system in erythrocytes and loss of this complex may contribute to acanthocytosis formation in McLeod syndrome.
Subject(s)
Amino Acid Transport Systems, Neutral/physiology , Cations, Divalent/blood , Erythrocytes/metabolism , Kell Blood-Group System/physiology , Abetalipoproteinemia/blood , Abetalipoproteinemia/genetics , Acanthocytes , Amino Acid Transport Systems, Neutral/blood , Amino Acid Transport Systems, Neutral/deficiency , Amino Acid Transport Systems, Neutral/genetics , Animals , Antiporters/blood , Calcium/blood , Calcium-Transporting ATPases/blood , Endothelin-3/pharmacology , Erythrocyte Volume , Genetic Diseases, X-Linked/blood , Genetic Diseases, X-Linked/genetics , Hemolysis/genetics , Homeostasis , Intermediate-Conductance Calcium-Activated Potassium Channels/blood , Ion Transport , Kell Blood-Group System/genetics , Magnesium/blood , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Multiprotein Complexes , Neuroacanthocytosis , Potassium/blood , Receptors, Endothelin/blood , Sodium/bloodABSTRACT
BACKGROUND: Kell and XK, two distinct red blood cell membrane proteins, are linked by a disulfide bond and form the Kell blood group complex. Kell surface antigens are expressed early during erythropoiesis but the onset of expression of XK which carries the Kx antigen is unknown. STUDY DESIGN AND METHODS: To determine whether Kell and XK are synchronously expressed, sorted human hematopoietic progenitor cells and mouse progenitor cells of defined lineage were studied. To determine the onset of expression, human marrow and cord blood cells were sorted into three subpopulations, representing stem, multipotent, and erythroid progenitor cells, and the expression of Kell and XK was determined by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. Mouse Kell and XK transcripts were determined by cDNA blotting of progenitor cells of defined lineage. RESULTS: By RT-PCR, human peripheral blood progenitor cells had weak expression of Kell and XK transcripts but FACS analysis did not detect surface antigens. Kell and XK transcripts are expressed in multipotent progenitor cells and these cells express Kell surface antigens. The expression of Kx antigen in progenitor cells was not determined owing to nonspecific reactions with the antibody. By cDNA blotting, mouse Kell expression was detected in bipotential megakaryocytes-erythroid cells and in colony-forming units-erythroid (CFU-E) and burst-forming units-erythroid (BFU-E), whereas XK was only detected in CFU-E and BFU-E. CONCLUSION: Both Kell and XK transcripts occur early during erythropoiesis; however, expression may not be coincident because, in mice, Kell transcripts, but not XK, occur in bipotential megakaryocytes-erythroid progenitor cells.