Degradation of serum amyloid A and apolipoproteins by serum proteases.
We have investigated the protease activity, present in human serum, that digests the serum amyloid A (SAA) protein. SAA radiolabeled with 125I was incubated at 37 degrees C with serum and plasma and analyzed for degradation products by alkaline urea-polyacrylamide gel electrophoresis and gel filtration chromatography. Serum initially digested SAA to intermediates of 3000-5000 in molecular weight, and these were further degraded to smaller peptides with prolonged incubation. SAA was not degraded by plasma anticoagulated with ethylenediaminetetraacetic acid (EDTA) or heparin. Recalcification of plasma anticoagulated with EDTA led to the generation of protease activity against SAA whereas EDTA plasma defibrinated with thrombin was inactive. We employed both nonselective and selective protease inhibitors and synthetic substrates for kallikrein and plasmin to further characterize the serum protease. These studies demonstrated that degradation of SAA is not directly attributable to enzymes involved in coagulation, kinin formation, or fibrinolysis, but the unidentified protease may be activated by one of the clotting factors. The specificity of the SAA degradation was demonstrated in experiments with three of the well-characterized apolipoproteins. Apolipoproteins A-I, C-I, and C-III-1, which also associate with the plasma high-density lipoproteins, were not degraded by serum although they were good substrates for purified thrombin and plasmin.
D000682:Amyloid / Q000378:metabolism*; D016632:Apolipoprotein A-I; D053302:Apolipoprotein C-I; D053305:Apolipoprotein C-III; D001053:Apolipoproteins / Q000378:metabolism*; D001056:Apolipoproteins C; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000097:blood*; D012697:Serine Endopeptidases; D000685:Serum Amyloid A Protein / Q000378:metabolism*
D000682:Amyloid; D016632:Apolipoprotein A-I; D053302:Apolipoprotein C-I; D053305:Apolipoprotein C-III; D001053:Apolipoproteins; D001056:Apolipoproteins C; D000685:Serum Amyloid A Protein; D006493:Heparin; D004492:Edetic Acid; D010450:Endopeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases