Protease nexin. Properties and a modified purification procedure.
The present paper describes chemical and functional properties of protease nexin, a serine protease inhibitor released from cultured human fibroblasts. It is shown that protease nexin is actually synthesized by fibroblasts and represents about 1% of their secreted protein. Analysis of the amino acid composition of purified protease nexin indicates that it is evolutionarily related to antithrombin III and heparin cofactor II. Protease nexin contains approximately 6% carbohydrate, with 2.3% amino sugar, 1.1% neutral sugar, and 3.0% sialic acid. The Mr calculated from equilibrium sedimentation analysis is 43,000. Protease nexin is a broad specificity inhibitor of trypsin-like serine proteases. It reacts rapidly with trypsin (kassoc = 4.2 +/- 0.4 X 10(6) M-1 s-1), thrombin (kassoc = 6.0 +/- 1.3 X 10(5) M-1 s-1), urokinase (kassoc = 1.5 +/- 0.1 X 10(5) M-1 s-1), and plasmin (kassoc = 1.3 +/- 0.1 X 10(5) M-1 s-1), and slowly inhibits Factor Xa and the gamma subunit of nerve growth factor but does not inhibit chymotrypsin-like proteases or leukocyte elastase. In the presence of heparin, protease nexin inhibits thrombin at a nearly diffusion-controlled rate. Two heparin affinity classes of protease nexin can be detected. The present characterization pertains to the fraction of protease nexin having the higher affinity for heparin. The low affinity material, which is the minor fraction, is lost during purification.
D000596:Amino Acids / Q000032:analysis; D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins / Q000378:metabolism*; D005347:Fibroblasts / Q000032:analysis; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008433:Mathematics; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000378:metabolism*; D058255:Protease Nexins; D011956:Receptors, Cell Surface; D013379:Substrate Specificity; D013917:Thrombin / Q000037:antagonists & inhibitors; D014357:Trypsin / Q000378:metabolism
D000596:Amino Acids; D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins; D011480:Protease Inhibitors; D058255:Protease Nexins; D011956:Receptors, Cell Surface; D006493:Heparin; D014357:Trypsin; D013917:Thrombin