• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of bone and mineral research: the official journal of the American Society for Bone and Mineral Research >Chondrogenic activity of the heparan sulfate proteoglycan perlecan maps to the N-terminal domain I.
【24h】

Chondrogenic activity of the heparan sulfate proteoglycan perlecan maps to the N-terminal domain I.

机译：硫酸乙酰肝素蛋白聚糖Perlecan的成软骨活性映射到N末端结构域I.

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

C3H10T1/2 cells differentiate along a chondrogenic pathway when plated onto the extracellular matrix (ECM) protein perlecan (Pln). To identify the region(s) within the large Pln molecule that provides a differentiation signal, recombinant Pln-sequence-based polypeptides representing distinct structural domains were assayed for their ability to promote chondrogenesis in C3H10T1/2 cells. Five distinct domains, along with structural variations, were tested. The N-terminal domain I was tested in two forms (IA and IB) that contain only heparan sulfate (HS) chains or both HS and chondroitin sulfate (CS) chains, respectively. A mutant form of domain I lacking attachment sites for both HS and CS (Pln I(mut)) was tested also. Other constructs consecutively designated Pln domains II, III(A-C), IV(A,B), and V(A,B) were used to complete the structure-function analysis. Cells plated onto Pln IA or Pln IB but no other domain rapidly assembled into cellular aggregates of 40-120 microm on average. Aggregate formation was dependent on the presence of glycosaminoglycan (GAG) chains, because Pln I-based polypeptides lacking GAG chains either by enzymatic removal or mutation of HS/CS attachment sites were inactive. Aggregates formed on GAG-bearing Pln IA stained with Alcian Blue and were recognized by antibodies to collagen type II and aggrecan but were not recognized by an antibody to collagen type X, a marker of chondrocyte hypertrophy. Collectively, these studies indicate that the GAG-bearing domain I of Pln provides a sufficient signal to trigger C3H10T1/2 cells to enter a chondrogenic differentiation pathway. Thus, this matrix proteoglycan (PG) found at sites of cartilage formation in vivo is likely to enhance early stage differentiation induced by soluble chondrogenic factors.
机译：C3H10T1 / 2细胞在接种到细胞外基质（ECM）蛋白Perlecan（Pln）上时会沿软骨形成途径分化。为了鉴定大Pln分子内提供分化信号的区域，测定了代表不同结构域的基于重组Pln序列的多肽在C3H10T1 / 2细胞中促进软骨形成的能力。测试了五个不同的域以及结构上的变化。 N端域I以两种形式（IA和IB）进行了测试，分别仅包含硫酸乙酰肝素（HS）链或HS和硫酸软骨素（CS）链。还测试了缺乏HS和CS附着位点的结构域I的突变形式（Pln I（mut））。依次指定为Pln域II，III（A-C），IV（A，B）和V（A，B）的其他构建体用于完成结构功能分析。细胞接种在Pln IA或Pln IB上，但没有其他域迅速组装成平均40-120微米的细胞聚集体。聚集体的形成取决于糖胺聚糖（GAG）链的存在，因为通过酶促去除或HS / CS附着位点的突变而缺乏GAG链的基于Pln I的多肽是无活性的。在载有GAG的Pln IA上形成的聚集体被Alcian Blue染色，并被II型胶原蛋白和聚集蛋白聚糖的抗体识别，但未被X型胶原蛋白（软骨细胞肥大的标志物）的抗体识别。总体而言，这些研究表明，Pln的带有GAG的结构域I提供了足以触发C3H10T1 / 2细胞进入软骨分化途径的信号。因此，在体内软骨形成部位发现的这种基质蛋白聚糖（PG）可能会增强由可溶性软骨形成因子诱导的早期分化。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号