A QUANTITATIVE-ANALYSIS OF THE INCORPORATION OF FIBULIN-1 INTO EXTRACELLULAR-MATRIX INDICATES THAT FIBRONECTIN ASSEMBLY IS REQUIRED
A QUANTITATIVE-ANALYSIS OF THE INCORPORATION OF FIBULIN-1 INTO EXTRACELLULAR-MATRIX INDICATES THAT FIBRONECTIN ASSEMBLY IS REQUIRED
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DOI：
10.1016/0945-053x(95)90004-7
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发表时间：
1995-02-01
期刊：
影响因子：
6.9
通讯作者：
ARGRAVES, WS
中科院分区：
文献类型：
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作者：
GODYNA, S;MANN, DM;ARGRAVES, WS
Fibulin-1 是一种细胞外基质糖蛋白，存在于疏松和致密结缔组织、弹性纤维和一些基底膜中。培养的细胞（例如成纤维细胞）将内源合成的或外源添加的 fibulin-1 组装成也含有纤连蛋白的基质原纤维。由于我们之前已证明 fibulin-1 与纤连蛋白结合（Balbona, K.、Tran, H.、Godyna, S.、Ingham, K.C.、Strickland, D.K. 和 Argraves, W.S. J. Biol. Chem. 267:20120-20125, 1992），因此我们试图研究 fibulin-1 掺入成纤维细胞中的情况细胞外基质，重点评估纤连蛋白在此过程中的潜在作用。在这项研究中，我们使用定量分析来测量 I-125-fibulin 与培养的成纤维细胞单层的结合。我们的结果表明，fibulin-1 掺入细胞层及其分配为去污剂可溶性和不溶性部分的动力学与纤连蛋白相似。发现纤连蛋白或纤连蛋白的 fibulin-1 结合域的抗体抑制 fibulin-1 掺入。无法将纤连蛋白组装到基质中的细胞系（例如 HT1080 或 PFHR-9）不会将 fibulin-1 整合到其细胞层中。然而，当 HT1080 细胞通过地塞米松处理诱导组装纤连蛋白时，它们随后获得了掺入 fibulin-1 的能力。此外，用抑制纤连蛋白组装的抗体处理培养的成纤维细胞可显着抑制 fibulin-1 掺入基质。当通过将细胞与外源纤连蛋白孵育不同时间长度而将增加量的纤连蛋白掺入细胞层时，还观察到fibulin-1掺入相应增加。综上所述，数据表明 fibulin-1 的掺入需要纤连蛋白组装，并表明依赖于基质中纤连蛋白的量。这些结果强调了纤连蛋白控制 fibulin-1 沉积到细胞外基质中的潜力，在细胞外基质中两种蛋白一致，并且可能对包含 fibulin-1 的基质结构（如基底膜或弹性纤维）的形成产生影响。
Fibulin-1 is an extracellular matrix glycoprotein found in both loose and dense connective tissues, elastic fibers and some basement membranes. Cultured cells such as fibroblasts assemble endogenously synthesized or exogenously added fibulin-1 into matrix fibrils that also contain fibronectin. Since we have previously shown that fibulin-1 binds to fibronectin (Balbona, K., Tran, H., Godyna, S., Ingham, K.C., Strickland, D.K. and Argraves, W.S. J. Biol. Chem. 267: 20120-20125, 1992), we sought to investigate fibulin-1 incorporation into fibroblast extracellular matrix with an emphasis on evaluating the potential role of fibronectin in the process. In this study, we have used quantitative assays to measure the binding of I-125-fibulin to monolayers of cultured fibroblasts. Our results show that the kinetics of fibulin-1 incorporation into the cell layer and its partitioning into detergent-soluble and -insoluble fractions were similar to those of fibronectin. It was found that antibodies to fibronectin or to the fibulin-1-binding domain of fibronectin-inhibited fibulin-1 incorporation. Cell lines that fail to assemble fibronectin into the matrix, such as HT1080 or PFHR-9, do not incorporate fibulin-1 into their cell layers. However, when HT1080 cells were induced to assemble fibronectin by treatment with dexamethasone, they subsequently acquired the ability to incorporate fibulin-1. Moreover, treatment of cultured fibroblasts with antibodies that inhibit fibronectin assembly significantly inhibit fibulin-1 incorporation into the matrix. When increased amounts of fibronectin were incorporated into cells layers by incubating the cells for varying lengths of time with exogenous fibronectin, a corresponding increase in fibulin-1 incorporation was also observed. Taken together, the data indicate that the incorporation of fibulin-1 requires fibronectin assembly and suggests a dependence on the amount of fibronectin in a matrix. These results highlight the potential of fibronectin to control the deposition of fibulin-1 into those extracellular matrices where both proteins coincide and may have implications in the formation of fibulin-1-containing matrix structures such as basement membranes or elastic fibers.