Isolation of platelet glycocalicin by affinity chromatography on thrombin-sepharose.
Platelet glycocalicin has been purified to homogeneity by a two step procedure involving affinity chromatography on WGA-Sepharose and then on thrombin-Sepharose using selective elution with heparin. The procedure is more rapid (3-4 days), more reproducible and gives about twice the yield (10 mg/40 units platelets) of the previous method (Okumura et al. J. Biol. Chem. 251, 5950-5955, 1976). The two preparations showed identical inhibition of aggregation of gel filtered platelets induced by thrombin and by ristocetin. Glycocalicin was cleaved in a controlled fashion by trypsin-Sepharose over 3hr to yield the macroglycopeptide and peptide "tail" fragments and there was no apparent further degradation with 18 hrs digestion.
D001792:Blood Platelets / Q000032:analysis; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification*; D006493:Heparin; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000302:isolation & purification*; D010974:Platelet Aggregation / Q000187:drug effects*; D019038:Platelet Glycoprotein GPIb-IX Complex; D012685:Sepharose; D013917:Thrombin; D013997:Time Factors
D006023:Glycoproteins; D008565:Membrane Proteins; D019038:Platelet Glycoprotein GPIb-IX Complex; C023592:glycocalicin; D006493:Heparin; D012685:Sepharose; D013917:Thrombin