Occurrence of platelet basic protein, a precursor of low affinity platelet factor 4 and beta-thromboglobulin, in human platelets and megakaryocytes.
beta-thromboglobulin antigen, a platelet-specific secreted protein, occurs in three forms: platelet basic protein, low affinity platelet factor 4, and beta-thromboglobulin. The combined level of beta-thromboglobulin antigen in megakaryocytes, measured by radioimmunoassay, was 13 +/- 7 micrograms/10(6) cells (SD, n = 6). The relative proportions of the three forms of beta-thromboglobulin antigen present within platelets and megakaryocytes were determined in cells lysed with trichloroacetic acid to minimize artifactual proteolysis. Samples were analyzed by isoelectric focusing in polyacrylamide gel with quantitative immunological detection on a nitrocellulose transfer of the gel. In platelets, the major species found was low affinity platelet factor 4 with precursor platelet basic protein as only 25% +/- 11% (SD, n = 16) of total beta-thromboglobulin antigen. In megakaryocytes partially purified both from normal bone marrow aspirates and from whole marrow specimens obtained after surgery, platelet basic protein was a higher proportion of beta-thromboglobulin antigen (49% +/- 13% SD, n = 11) than was the case in platelets. beta-thromboglobulin itself was never detected under the conditions of cell lysis used. Our results suggest that platelet basic protein is synthesized in megakaryocytes and that its cleavage is associated with an earlier stage of cell development than simply maturation to platelets. Further support for the precursor status of platelet basic protein was found in the expression of predominantly this antigenic form in a human erythroleukemia cell line.
Holt
J C
JC
Thrombosis Research Center, Temple University, Philadelphia, Pennsylvania 19140.
Rabellino
E M
EM
Gewirtz
A M
AM
Gunkel
L M
LM
Rucinski
B
B
Niewiarowski
S
S
D016824:Antigens, Human Platelet; D001779:Blood Coagulation Factors / Q000032:analysis* / Q000302:isolation & purification; D001792:Blood Platelets / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D018925:Chemokines; D006801:Humans; D039661:Integrin beta3; D007519:Isoantigens / Q000032:analysis* / Q000302:isolation & purification; D004915:Leukemia, Erythroblastic, Acute / Q000032:analysis / Q000097:blood / Q000276:immunology; D008533:Megakaryocytes / Q000032:analysis* / Q000276:immunology; D010455:Peptides; D011498:Protein Precursors / Q000097:blood* / Q000302:isolation & purification; D011506:Proteins / Q000302:isolation & purification; D014407:Tumor Cells, Cultured; D001620:beta-Thromboglobulin / Q000276:immunology*
D016824:Antigens, Human Platelet; D001779:Blood Coagulation Factors; D018925:Chemokines; C061248:ITGB3 protein, human; D039661:Integrin beta3; D007519:Isoantigens; C061615:PPBP protein, human; D010455:Peptides; D011498:Protein Precursors; D011506:Proteins; D001620:beta-Thromboglobulin; C009862:connective tissue-activating peptide; C018329:low affinity platelet factor 4