Identification of cytokeratin 1 as a binding protein and presentation receptor for kininogens on endothelial cells
Identification of cytokeratin 1 as a binding protein and presentation receptor for kininogens on endothelial cells
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DOI：
10.1073/pnas.95.7.3615
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发表时间：
1998-03-31
影响因子：
11.1
通讯作者：
Schmaier, AH
中科院分区：
文献类型：
--
作者：
Hasan, AAK;Zisman, T;Schmaier, AH
在内皮细胞上鉴定了一种激肽原结合蛋白（一种假定的受体），通过生物素-高分子量激肽原（HK）亲和柱分离了一种54-kDa蛋白，在胰蛋白酶消化酶的氨基末端测序中，该蛋白被鉴定为细胞角蛋白1，针对细胞角蛋白1的多种抗体与内皮细胞裂解物免疫印迹中的54-kDa条带反应，在激光扫描中：共聚焦显微镜下，在内皮细胞表面发现了细胞角蛋白1抗原，流式细胞术也在内皮细胞膜上检测到细胞角蛋白1抗原，此外，针对细胞角蛋白1独特序列的抗肽抗体也特异性地结合于未透化的内皮细胞。HK和低分子量的激肽原（而非因子XII）可阻断HK与细胞角蛋白的结合，HK结构域3、4和5上的细胞结合区的多肽可阻断HK与细胞角蛋白的结合。gC 1 qR和可溶性尿激酶样纤溶酶原激活物受体也抑制生物素-HK与细胞角蛋白的结合。这些研究证实了细胞角蛋白1作为激肽原结合蛋白的新功能。
A kininogen binding protein(s), a putative receptor, was identified on endothelial cells, A 54-kDa protein was isolated by a biotin-high molecular mass kininogen (HK) affinity column that, on aminoterminal sequencing of tryptic digests, was identified as cytokeratin 1, Multiple antibodies directed to cytokeratin 1 reacted with a 54-kDa band on immunoblot of lysates of endothelial cells, On laser scanning: confocal microscopy, cytokeratin 1 antigen was found on the surface of endothelial tells, Cytokeratin 1 antigen also was detected on. endothelial cell membranes by flow cytometry, Moreover, an antipeptide antibody to a sequence unique to cytokeratin 1 also specifically bound to nonpermeabilized endothelial cells. Biotin-HK specifically bound to cytokeratin only in the presence of Zn2+, and cytokeratin blocked biotin-HK binding to endothelial cells, Further, HK and low molecular mass kininogen, but not factor XII, blocked biotin-HK binding to cytokeratin, and pep tides of each cell binding region of HK on domains 3,4, and 5 blocked biotin-HK binding to cytokeratin. gC1qR and soluble urokinase-like plasminogen activator receptor also inhibited biotin-HK binding to cytokeratin, These investigations identify a new function for cytokeratin 1 as a kininogen binding protein, Cytokeratins, members of the family of intermediate filament proteins, may represent a new class of receptors.