Dermatopontin Promotes Epidermal Keratinocyte Adhesion via α3β1 Integrin and a Proteoglycan Receptor
Dermatopontin Promotes Epidermal Keratinocyte Adhesion via α3β1 Integrin and a Proteoglycan Receptor
复制标题
DOI：
10.1021/bi901066f
复制
发表时间：
2010-01-12
期刊：
影响因子：
2.9
通讯作者：
Fujiwara, Sakuhei
中科院分区：
文献类型：
--
作者：
Okamoto, Osamu;Hozumi, Kentaro;Fujiwara, Sakuhei
皮桥蛋白是一种最初从牛真皮中提纯的细胞外基质成分，可促进人表皮角质形成细胞系(HaCaT细胞)的细胞黏附。HaCaT细胞散布在皮桥蛋白上，形成肌动蛋白纤维。EDTA和肝素均可抑制HaCaT细胞与皮桥蛋白的黏附，并部分由α3β1整合素介导。一种合成肽(DP-4，PHGQVVVAVRS；牛皮桥蛋白残基33-43)特异性地抑制细胞与皮桥蛋白的黏附，当DP-4肽包被在井上时，它以剂量依赖的方式促进细胞黏附。DP-4肽的活性核心序列被定位为八个氨基酸序列(GQVVVAVR)。这些结果表明，皮桥蛋白是一种新的表皮细胞黏附分子，提示DP-4序列在皮桥蛋白的细胞黏附活性中起关键作用。肝素可显著抑制细胞与DP-4的黏附。用肝素酶1处理HaCaT细胞时，细胞不能与DP-4黏附，但软骨素酶ABC处理不影响黏附活性。DP-4与生物素标记的肝素发生特异性相互作用，这种作用被未标记的肝素抑制。DP-4多肽显著促进过表达Syndecans的细胞的黏附，Syndecan与DP-4多肽亲和柱结合。这些结果表明，HaCaT细胞通过α3β1整合素和硫酸乙酰肝素蛋白多糖类型的受体与皮桥蛋白黏附，这可能是一种Syndecan。我们认为，皮桥蛋白是一种多功能的表皮细胞黏附分子。
Dermatopontin, an extracellular matrix component Initially purified from bovine dermis, promoted cell adhesion Of the human epidermal keratinocyte cell line (HaCaT cells). HaCaT cells spread on dermatopontin and formed actin fibers. Adhesion of HaCaT cells to dermatopontin was inhibited by both EDTA and heparin and was mediated in part by alpha 3 beta 1 Integrin. A synthetic peptide (DP-4, PHGQVVVAVRS; bovine dermatopontin residues 33-43) specifically inhibited adhesion of cells to dermatopontin, and when the DP-4 peptide was coated on the well, it promoted cell adhesion in a dose-dependent manner. An active core Sequence of the DP-4 peptide was localized to an eight-amino acid sequence (GQVVVAVR). These results indicate that dermatopontin is a novel epidermal cell adhesion molecule and suggest that the DP-4 sequence is critical for the cell adhesive activity of dermatopontin. Adhesion of cells to DP-4 was strongly inhibited by heparin. When HaCaT cells were treated with heparitinase 1, the cells failed to adhere to DP-4 but chondroitinase ABC treatment did not Influence the adhesion activity. DP-4 specifically interacted with biotinylated heparin, and this interaction was inhibited by unlabeled heparin. DP-4 peptide significantly promoted the adhesion of cells overexpressing syndecans, and syndecan bound to a DP-4 peptide affinity column. These results suggest that HaCaT cells adhere to dermatopontin through alpha 3 beta 1 integrin and a heparan sulfate proteoglycan-type receptor, which is likely a syndecan. We conclude that dermatopontin plays a role as a multifunctional adhesion molecule for epidermal cells.