Activated protein C is not regulated by alpha-2-macroglobulin in plasma.
Alpha-2-macroglobulin (a2M) is a wide spectrum plasma inhibitor which functions by a unique mechanism and is a secondary inhibitor of coagulation and fibrinolytic enzymes. Human activated protein C (APC) is the central enzyme of a major regulatory system of coagulation and fibrinolysis. APC is primarily regulated (inhibited) by a specific plasma inhibitor. We undertook this study to investigate the role of a2M as a secondary inhibitor of APC. APC did not interact with a2M by any of the known mechanisms of interaction. APC failed to bind and form the classic proteinase-a2M complex as seen with similar serine proteases, thrombin and trypsin. APC also failed to cleave the a2M molecule. Experiments, using purified APC and either purified a2M or plasma, failed to demonstrate APC binding to a2M in gel filtration chromatography. No enzymatic activity of APC or radiolabeled APC was demonstrated in the a2M peak. Using an immuno-enzymatic assay (Harpel, J Biol Chem 260:4257, 1985) for an a2M and enzyme complex, the amount of APC bound to a2M was less than 3% of the added APC (non-specific binding only); whereas in similar experiments with thrombin, 75-86% of the added trypsin or thrombin bound. These data demonstrate that APC is one of a small number of unique serine proteases that do not interact with a2M. The absence of sequence homology between the a2M 'bait region' and the APC substrate cleavage sequences appear to be the reason APC is not inhibited by a2M.
Marlar
R A
RA
Department of Pediatrics, University of Colorado Health Sciences Center, Denver 80262.
Kressin
D C
DC
D002850:Chromatography, Gel; D004789:Enzyme Activation; D006801:Humans; D007124:Immunoenzyme Techniques; D007531:Isoflurophate; D011486:Protein C / Q000502:physiology*; D000511:alpha-Macroglobulins / Q000502:physiology*
D011486:Protein C; D000511:alpha-Macroglobulins; D007531:Isoflurophate