Binding properties of Pyrularia thionin and Naja naja kaouthia cardiotoxin to human and animal erythrocytes and to murine P388 cells.
Pyrularia thionin and snake venom cardiotoxin are strongly basic peptides which induce hemolysis, depolarization of muscle cells and activation of endogenous phospholipase A2. An earlier study of the hemolysis reaction indicated that the two peptides bind to and compete for the same site on human erythrocytes. A recent study examined the hemolysis induced by both peptides as the phosphate and Ca2+ content of the reaction mixture was varied. The results of the recent study (VERNON, L. P. and ROGERS, A., Toxicon 30, 701-709) agree with this companion study on the binding of 125I-labeled pyrularia thionin and cardiotoxin to erythrocytes under the same conditions. Added phosphate ion at 5 mM and removal of membrane-bound Ca2+ by treatment with 10 mM EGTA make more binding sites of the same affinity available to both peptides, which are shown to bind in a competitive fashion to the same site. Addition of 10 mM Ca2+ to the medium decreases peptide binding due to competitive binding of Ca2+ to the same site on the membrane. For human erythrocytes the number of binding sites/cell for the thionin ranged from 0.7 to 1.7 x 10(5) and for cardiotoxin from 0.82 to 1.6 x 10(5). The calculated dissociation constants (Kd) from the Scatchard plots ranged from 0.43 to 1.1 microM for the thionin and from 0.40 to 0.98 microM for the cardiotoxin. The binding sites for thionin and cardiotoxin with sheep erythrocytes were 1.7 and 2.0 x 10(4) sites/cell, respectively, and both cow and horse erythrocytes demonstrated 2.7 x 10(4) sites/cell for the thionin. Binding studies with murine P388 cells showed 7.0 and 9.5 x 10(6) sites per cell for Pyrularia thionin and cardiotoxin, respectively.
Vernon
L P
LP
Chemistry Department, Brigham Young University, Provo, UT 84602.
Rogers
A
A
D000818:Animals; D023181:Antimicrobial Cationic Peptides; D001665:Binding Sites; D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D004179:Cobra Cardiotoxin Proteins / Q000378:metabolism*; D003603:Cytotoxins / Q000378:metabolism*; D004533:Egtazic Acid / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007941:Leukemia P388 / Q000378:metabolism*; D051379:Mice; D010940:Plant Proteins / Q000378:metabolism*; D010947:Plants, Toxic; D014407:Tumor Cells, Cultured
D023181:Antimicrobial Cationic Peptides; D004179:Cobra Cardiotoxin Proteins; D003603:Cytotoxins; D010940:Plant Proteins; C044942:THI1 protein, Pyrularia pubera; D004533:Egtazic Acid; D002118:Calcium