Novel structurally altered P(2X1) receptor is preferentially activated by adenosine diphosphate in platelets and megakaryocytic cells.
Experimental and clinical data suggest the presence of multiple types of adenosine diphosphate (ADP) receptors, one coupled to ligand-gated cation channels (P(2X)) and others coupled to G-protein-coupled (P(2Y)) receptors. This report identifies cDNA for a structurally altered P(2X1)-like receptor in megakaryocytic cell lines (Dami and CMK 11-5) and platelets that, when transfected into nonresponsive 1321 cells, confers a specific sensitivity to ADP with the pharmacologic rank order of ADP > > ATP > > > alpha,beta-methylene-ATP as measured by Ca(++) influx. This receptor (P(2X1del)) contains a deletion of 17 amino acids (PALLREAENFTLFIKNS) that includes an NFT consensus sequence for N-linked glycosylation. Glycosylated forms of the P(2X1del) and P(2X1wt) receptors were indistinguishable electrophoretically by Western blot or by immunoprecipitation using available antihuman and antirat antibodies. These results indicate that the expression of the P(2X1del) receptor results in an influx of Ca(++) induced by ADP. Expression of P(2X1del) receptor homomeric subunits is sufficient to express a receptor preferentially activated by ADP and suggests that this altered form, alone or in combination with P(2X1wt) receptors, is a component of an ADP-activated ion channel.
Greco
N J
NJ
Platelet Biology and the Product Development Departments, American Red Cross, Rockville, MD, USA. greco@usa.redcross.org
Tonon
G
G
Chen
W
W
Luo
X
X
Dalal
R
R
Jamieson
G A
GA
D000244:Adenosine Diphosphate / Q000494:pharmacology*; D001792:Blood Platelets / Q000378:metabolism*; D020013:Calcium Signaling / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008533:Megakaryocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011983:Receptors, Purinergic / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D018048:Receptors, Purinergic P2 / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D058469:Receptors, Purinergic P2X; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D011983:Receptors, Purinergic; D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D000244:Adenosine Diphosphate