| Summary: | 碩士 === 國立中正大學 === 化學工程研究所 === 105 === Atherosclerosis is considered as a chronic inflammatory disease. Low density lipoprotein (LDL) has been shown as a risk factor for atherosclerosis. Once oxidized, oxidized LDL (oxLDL) is toxic to a various types of cells, contributing to the atherosclerotic plaque formation through induction of endothelial cell activation and dysfunction, foam cell formation, and smooth muscle cell proliferation. C-reactive protein (CRP) is not only an inflammatory marker, but also directly involves in the pathogenesis of atherothrombosis. Previously, we found that keratin-1 (KRT1) binds to CRP and mediates the CRP effect on reduction of nitric oxide release. To further elucidate the interaction of KRT1 with CRP and oxLDL. The KRT1 protein was expressed in E. coli and purified using Ni2+-column. The interaction of KRT1 with CRP and oxLDL was assayed by the enzyme-linked immunosorbent assay (ELISA). Our results showed that the binding pattern of KRT1 to CRP was dependent on the KRT1 conformation obtained from the renaturing process. KRT1 renatured with Tween 20 showed a calcium-independent binding to CRP and the presence of phosphocholine (PC) did not inhibit their interaction. In contrast, KRT1 recovered in the absence of Tween 20 depended on calcium to bind CRP and addition of PC inhibited their interaction. We also confirmed that KRT1 bound to oxLDL. The binding of KRT1 to CRP and oxLDL strongly suggests the involvement of KRT1 in the process of atherosclerosis.
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