Antigenic and antiheparin properties of human platelet factor 4 (PF4)

Blood. 1975 Apr;45(4):537-50.

Abstract

Platelet factor 4 (PF4, a heparin-neutralizing protein) was isolated from washed human platelets. It was found to be homogenous by SDS-polyacrylamide gel electrophoresis, immunodiffusion, and immunoelectrophoresis, when tested with monospecific antibody produced in rabbits. PF4 is a heat-stable protein, but its antiheparin activity and antigenicity are destroyed by trypsin. The molecular weight of PF4 as calculated by amino acid analysis is approximately 8000 and by SDS-polyacrylamide gel electrophoresis with beta-mercaptoethanol, 7100 daltons. PF4 migrated to the cathode at pH 8.6. The interaction of PF4 with heparin resulted in the formation of a complex which migrated to the anode, as tested by immunoelectrophoresis. Incubation of purified PF4 with its antibody at 37 degrees C resulted in a loss of antiheparin activity. The presence of antiheparin activity and of PG4 antigen in material released during platelet aggregation by various agents and at various stages of the preparative procedure closely correlated. It has been concluded that PF4 antigen and antiheparin activity are two properties of the same protein. Comparison of human and pig PF4 revealed significant biochemical and antigenic differences.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Antigens*
  • Blood Coagulation Factors / immunology*
  • Blood Platelets*
  • Blood Proteins / analysis
  • Chromatography, DEAE-Cellulose
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Heparin / pharmacology
  • Heparin Antagonists*
  • Humans
  • Immune Sera
  • Immunodiffusion
  • Immunoelectrophoresis
  • Molecular Weight
  • Platelet Aggregation
  • Polyethylene Glycols
  • Protamines
  • Rabbits / immunology
  • Serum Albumin
  • Sodium Dodecyl Sulfate
  • Swine / immunology
  • Trypsin / pharmacology

Substances

  • Amino Acids
  • Antigens
  • Blood Coagulation Factors
  • Blood Proteins
  • Heparin Antagonists
  • Immune Sera
  • Protamines
  • Serum Albumin
  • Sodium Dodecyl Sulfate
  • Polyethylene Glycols
  • Heparin
  • Trypsin