Human platelet basic protein associated with antiheparin and mitogenic activities: purification and partial characterization.
Platelet basic protein (PBP) (isoelectric point, 10.0-10.5; apparent Mr, 11,000-15,000) has been purified to homogeneity from material secreted by fresh human platelets after stimulation by thrombin. The purification, using preparative isoelectric focusing and chromatography on heparin-Sepharose, yielded two additional peptides with antiheparin activity that were immunologically identical with PBP: low-affinity platelet factor 4 and beta-thromboglubulin. The purity of the peptides was confirmed by immuoelectrophoresis and by NH2-terminal amino acid analysis. Dansyl chloride-treated PBP yielded a single dansylated amino acid residue (glycine). By using a specific radioimmunoassay it was shown that 10(9) human platelets contain 2-3 microgram of PBP which can be released in response to specific stimulation. PBP is associated with mitogenic activity as assayed in Swiss 3T3 mouse cells cultured in low-serum (0.4-1.5%) medium at levels of about 1 ng/ml and saturating at 10-40 ng/ml. The biological activity of different PBP preparations was variable, presumably due to inhibition by the varying amounts of ampholytes that interfered with the mitogenic activity of the peptide. Mitogenic activity was eluted from NaDodSO4/polyacrylamide gels and shown to comigrate with immunoreactive material and with conventional marker proteins of 14,000-17,000 daltons or with histones of 11,000-15,000 daltons. Evidence is presented that PBP is different from cationic platelet-derived growth factor which has an apparent Mr of 30,000.
D000595:Amino Acid Sequence; D000818:Animals; D001681:Biological Assay; D001779:Blood Coagulation Factors / Q000302:isolation & purification*; D001792:Blood Platelets / Q000032:analysis*; D002478:Cells, Cultured; D006133:Growth Substances / Q000097:blood*; D006801:Humans; D007526:Isoelectric Point; D051379:Mice; D008934:Mitogens / Q000097:blood*; D010978:Platelet Factor 4 / Q000276:immunology / Q000302:isolation & purification*
D001779:Blood Coagulation Factors; D006133:Growth Substances; D008934:Mitogens; D010978:Platelet Factor 4