Characterization of human platelet basic protein, a precursor form of low-affinity platelet factor 4 and beta-thromboglobulin.
Platelet basic protein (PBP) was purified from the supernatant of thrombin-stimulated, washed human platelets by ion-exchange, affinity, molecular sieve, and high-performance liquid chromatography (HPLC). The NH2-terminal amino acid sequence was determined by automated Edman degradation, revealing 9 unique residues followed by 10 residues of the established low-affinity platelet factor 4/beta-thromboglobulin (LA-PF4/beta TG) sequence. Among the nine were three basic residues, accounting for the high isoelectric point of PBP. Additional evidence for precursor status includes the immunological cross-reactivity of all three species and the ability of plasmin and trypsin to produce from PBP a species resembling beta TG in charge, hydrophobicity, and size. Tryptic peptide maps of PBP and LA-PF4 obtained by reverse-phase HPLC were very similar, and from each protein, a peptide was isolated which showed the amino acid composition predicted for the COOH-terminal tryptic peptide of beta TG. Normal platelets contained predominantly LA-PF4, with PBP ranging from 10% to 30% of total beta TG antigen. This was true even when fresh platelets were lysed with trichloroacetic acid in order to provide the most complete and rapid inhibition of proteolytic activity. beta TG itself was never detected in this situation or in the release supernatant of stimulated platelets, and only rarely in unprotected lysates. In agreement with earlier results, crude preparations of PBP were mitogenic for 3T3 cells, but highly purified preparations of PBP and LA-PF4 were free of this activity.
J C
JC
Holt
M E
ME
Harris
A M
AM
Holt
E
E
Lange
A
A
Henschen
S
S
Niewiarowski
D000595:Amino Acid Sequence; D001609:Beta-Globulins / Q000096:biosynthesis*; D001792:Blood Platelets / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007106:Immune Sera; D007525:Isoelectric Focusing; D008934:Mitogens; D010446:Peptide Fragments / Q000032:analysis; D010978:Platelet Factor 4 / Q000096:biosynthesis*; D011498:Protein Precursors / Q000302:isolation & purification*; D011863:Radioimmunoassay; D001620:beta-Thromboglobulin / Q000096:biosynthesis*
D001609:Beta-Globulins; D007106:Immune Sera; D008934:Mitogens; D010446:Peptide Fragments; D011498:Protein Precursors; D001620:beta-Thromboglobulin; D010978:Platelet Factor 4