Endogenous heparinase-sensitive anticoagulant activity in human plasma.
In this paper we show that an anticoagulant activity, which we measure by thrombin time, appears in human plasma after its exhaustive proteolytic digestion. This activity is extremely heat stable, it is resistant to chondroitin ABC lyase (E.C.4.2.2.4) and heparan sulfate lyase (E.C.4.2.2.8), it is sensitive to heparin lyase (E.C.4.2.2.7) and to nitrous acid treatment: we suggest that it can be identified as authentic heparin. The amount present in 1 ml of plasma of healthy subjects corresponds to 0.1-0.2 I.U. of standard heparin (150 I.U./mg). Proteolytically digested human plasma was submitted to ion-exchange chromatography on DEAE-Sephacel and the anticoagulant activity in the fractions eluted at the different molarities of NaCl was measured by thrombin time. This analysis shows that the anticoagulant activity elutes at very low ionic strength. The possibility that interactions of the endogenous heparin with proteins or protein fragments are responsible for the difficulty in isolating heparin from human plasma is discussed.
Cavari
S
S
Institute of General Pathology, University of Firenze, Italy.
Stramaccia
L
L
Vannucchi
S
S
D002852:Chromatography, Ion Exchange; D002918:Chymotrypsin; D017364:Collagenases; D004591:Electrophoresis, Polyacrylamide Gel; D006493:Heparin / Q000032:analysis*; D006801:Humans; D010206:Papain; D010434:Pepsin A; D010949:Plasma / Q000737:chemistry*; D014357:Trypsin
D006493:Heparin; D002918:Chymotrypsin; D014357:Trypsin; D010206:Papain; D010434:Pepsin A; D017364:Collagenases