Expression of proteoglycan core proteins in human bone marrow stroma.
Expression of proteoglycan core proteins in human bone marrow stroma.
复制标题
人骨髓基质中蛋白多糖核心蛋白的表达。
DOI：
10.1042/bj3430663
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
G. David
中科院分区：
文献类型：
--
作者：
K. Schofield;J. Gallagher;G. David
硫酸乙酰肝素蛋白多糖(HSPGs)存在于骨髓基质细胞表面和细胞外基质(ECM)中，对造血干/祖细胞的黏附和生长具有重要的调控作用。含有硫酸乙酰肝素链的两类主要蛋白多糖是Syndecan和Glypcan家族的成员。在这项研究中，我们鉴定了长期培养形成的正常人骨髓基质中存在的主要表面膜和基质相关的热休克蛋白。从贴壁的基质层中提取蛋白多糖，用肝素酶和软骨素酶ABC处理。用针对syndecans-1-4，glypcan-1和ECM HSPG，perlecan的抗体进行Western blotting检测核心蛋白。Northern印迹和逆转录聚合酶链式反应检测基质细胞在RNA水平的表达。Glypcan-1、Syndecan-3和Syndecan-4是主要的细胞膜HSPG种类，Perlecan是主要的ECM蛋白多糖。未见Syndecan-1蛋白表达。Syndecan-3主要以突变体或加工后的50-55 kDa核心蛋白的形式表达，少量表达为125 kDa的特征核心蛋白。这些结果表明，骨髓基质细胞表面存在Syndecan-3、Syndecan-4和Glypcan-1，与ECM中的Perlecan一起，可能为造血干和祖细胞的维持和发育创造了正确的基质‘生态位’。作为主要基质HSPG的Syndecan-3变异形式的检测表明了Syndecan-3在造血中的特殊作用。
Heparan sulphate proteoglycans (HSPGs) present on the surface of bone marrow stromal cells and in the extracellular matrix (ECM) have important roles in the control of adhesion and growth of haemopoietic stem and progenitor cells. The two main groups of proteoglycans which contain heparan sulphate chains are members of the syndecan and glypican families. In this study we have identified the main surface membrane and matrix-associated HSPGs present in normal human bone marrow stroma formed in long-term culture. Proteoglycans were extracted from the adherent stromal layers and treated with heparitinase and chondroitinase ABC. The core proteins were detected by Western blotting using antibodies directed against syndecans-1-4, glypican-1 and the ECM HSPG, perlecan. Stromal cell expression at the RNA level was detected by Northern blotting and by reverse transcription PCR. Glypican-1, syndecan-3 and syndecan-4 were the major cell-membrane HSPG species and perlecan was the major ECM proteoglycan. There was no evidence for expression of syndecan-1 protein. Syndecan-3 was expressed mainly as a variant or processed 50-55 kDa core protein and in lower amounts as the characteristic 125 kDa core protein. These results suggest that syndecan-3, syndecan-4 and glypican-1 present on the surface of marrow stromal cells, together with perlecan in the ECM, may be responsible for creating the correct stromal 'niche' for the maintenance and development of haemopoietic stem and progenitor cells. The detection of a variant form of syndecan-3 as a major stromal HSPG suggests a specific role for this syndecan in haemopoiesis.