G1 Domain of Versican Regulates Hyaluronan Organization and the Phenotype of Cultured Human Dermal Fibroblasts.
G1 Domain of Versican Regulates Hyaluronan Organization and the Phenotype of Cultured Human Dermal Fibroblasts.
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DOI：
10.1369/0022155416643913
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发表时间：
2016-06
期刊：
影响因子：
--
通讯作者：
Wight TN
中科院分区：
文献类型：
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作者：
Merrilees MJ;Zuo N;Evanko SP;Day AJ;Wight TN
多功能蛋白聚糖的变体对细胞和组织表型具有广泛的影响，影响增殖、粘附、细胞周围基质组成和弹性蛋白生成。多功能蛋白聚糖的G1结构域，其中包含两个连接模块，结合透明质酸（HA），可能是中央的这些影响。重组人G1（rhG 1）与N-末端8个氨基酸的组氨酸（His）标签，产生于烟草本塞姆氏，施加到培养的皮肤成纤维细胞，增殖和细胞周HA组织的影响确定。rhG 1定位于细胞表面HA的单个链，其聚集成类似HA电缆的结构。在单个链和聚集链上，附着的rhG 1的间距相似（约120 nm），表明rhG 1分子之间存在相互作用。内源性V0/V1，存在于连接rhG 1之间的HA上，不能阻止电缆形成，而单独使用V0/V1治疗，也与HA结合，不能诱导电缆形成。rhG 1单次处理可长时间抑制细胞增殖。用rhG 1处理细胞4周导致细长的、分化的α肌动蛋白阳性成纤维细胞的浓缩层，rhG 1定位于细胞表面，以及包括胶原蛋白和弹性蛋白的致密细胞外基质。这些结果表明，多功能蛋白聚糖的G1结构域可以调节细胞周HA的组织和影响表型。
Variants of versican have wide-ranging effects on cell and tissue phenotype, impacting proliferation, adhesion, pericellular matrix composition, and elastogenesis. The G1 domain of versican, which contains two Link modules that bind to hyaluronan (HA), may be central to these effects. Recombinant human G1 (rhG1) with an N-terminal 8 amino acid histidine (His) tag, produced in Nicotiana benthamiana, was applied to cultures of dermal fibroblasts, and effects on proliferation and pericellular HA organization determined. rhG1 located to individual strands of cell surface HA which aggregated into structures resembling HA cables. On both individual and aggregated strands, the spacing of attached rhG1 was similar (~120 nm), suggesting interaction between rhG1 molecules. Endogenous V0/V1, present on HA between attached rhG1, did not prevent cable formation, while treatment with V0/V1 alone, which also bound to HA, did not induce cables. A single treatment with rhG1 suppressed cell proliferation for an extended period. Treating cells for 4 weeks with rhG1 resulted in condensed layers of elongated, differentiated α actin-positive fibroblasts, with rhG1 localized to cell surfaces, and a compact extracellular matrix including both collagen and elastin. These results demonstrate that the G1 domain of versican can regulate the organization of pericellular HA and affect phenotype.