Characterization of platelet agglutinating protein p37 purified from the plasma of a patient with thrombotic thrombocytopenic purpura.
We have previously reported the purification of a 37 KDa platelet agglutinating protein (PAP p37) from the plasma of a patient with thrombotic thrombocytopenic purpura and have shown that it is present in a subset of TTP patients, but absent in normal subjects. In this study, we would like to report some of the physico-chemical and immunological properties of this protein. The native molecular weight of PAP p37 from gel filtration was found to be 36,000, which is in agreement with denatured molecular weight (37,000), determined by SDS--polyacrylamide gel electrophoresis under both reducing and non-reducing conditions. The values of Stoke's radius (25A), diffusion coefficient (8.59 x 10(-7)cm2/s) and frictional ratio (1.13), determined by molecular sieve chromatography, suggest that the native protein is compact and globular. The purified protein has an S20,w of 3.5s. Preliminary carbohydrate analysis suggested that p37 is a glycoprotein and contained about 11% neutral sugars and 6.6% sialic acid. Amino acid analysis indicated that the protein is relatively rich in aspartate and serine and has low cysteine, methionine and tryptophan contents. In dot immunobinding ELISA assay, PAP p37 did not react with antibodies to thrombospondin, fibrinogen, fibronectin, plasminogen and von Willebrand factor. Our results suggest that PAP p37 is a single polypeptide compact and globular glycoprotein and is immunologically not related to the aforementioned proteins.
Siddiqui
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FA
Division of Hematology/Oncology, Sylvester Cancer Center, University of Miami School of Medicine, Florida.
Lian
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EC
D000596:Amino Acids / Q000032:analysis; D001792:Blood Platelets / Q000502:physiology*; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002241:Carbohydrates / Q000032:analysis; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D010974:Platelet Aggregation / Q000187:drug effects; D011480:Protease Inhibitors / Q000494:pharmacology*; D011487:Protein Conformation; D011697:Purpura, Thrombotic Thrombocytopenic / Q000097:blood*; D012794:Sialic Acids / Q000032:analysis
D000596:Amino Acids; D001798:Blood Proteins; D002241:Carbohydrates; D011480:Protease Inhibitors; D012794:Sialic Acids; C046935:platelet agglutinating protein, thrombotic thrombocytopenic purpura