Characterization of serum amyloid P component from human aortic atherosclerotic lesions

Abstract

Serum amyloid P component (SAP) is a glycoprotein in human plasma. We recently showed the localization of SAP in human atherosclerotic lesions by immunohistochemical staining. In this study, the presence of SAP in atherosclerotic lesions was confirmed, and the biochemical character of SAP in atherosclerotic intima was investigated and compared with that of native SAP. Atherosclerotic intima was sequentially extracted with 2 mmol/L CaCl2-Tris-buffered saline (TBS), 10 mmol/L EDTA-TBS, 3 mol/L guanidine-TBS, and collagenase digestion. The character of SAP in each extract was studied with double immunodiffusion, electroimmunoassay, crossed immunoelectrophoresis, and Western immunoblotting. The total amount of SAP in atherosclerotic intima was 190 +/- 64 micrograms/g wet tissue with an SAP-albumin ratio of 1:22.7, which is 44 times higher than the relative plasma ratio of 1:1000. This suggests that SAP is specifically localized in atherosclerotic lesions. SAP from the intima was indistinguishable from plasma or purified SAP with respect to immunological character and molecular weight. However, electrophoretic mobility and the binding of SAP to atherosclerotic intima appeared heterogeneous. Of total extractable SAP, about 43% appeared in the CaCl2-TBS fraction, 25% in the EDTA-TBS fraction, and 32% in the collagenase digestion fraction. SAP is one of the two pentraxins in human plasma; the other is C-reactive protein, which has also been reported to locate in atherosclerotic lesions. Our findings suggest a role for SAP in atherogenesis and encourage efforts to determine more precisely the physiological contributions of the pentraxin family to the development of atherosclerosis.