Okuda H, Kamada A
Department of Biochemistry, Osaka Dental University, Japan.
J Osaka Dent Univ. 1999 Oct;33(2):59-64.
We investigated the characteristics of cell-membrane and extracellular proteoglycans in the monkey submandibular gland using enzymatic digestion and Western blot analysis. Extracellular proteoglycans and cell-membrane proteoglycans were extracted from the phosphate-buffered saline (PBS) soluble fraction and microsomal fraction of submandibular glands, respectively, and purified partially by ion exchange chromatography. Cellulose acetate membrane electrophoresis and immunoblotting with 3G10 monoclonal antibody or 6B6 monoclonal antibody showed that the extracellular proteoglycans contain several heparan sulfate proteoglycans (117, 67, 47 and 35 kDa core proteins) and a dermatan sulfate proteoglycan (45-50 kDa core protein), while the cell-membrane proteoglycans contain only a heparan sulfate proteoglycan with 69 kDa core protein. These results indicate that the several extracellular heparan sulfate proteoglycans may be released ectodomains of the cell-membrane proteoglycans from cell surface. It is suggested that the cell-membrane and extracellular proteoglycans may regulate the function of growth factors and the microenvironment in the normal submandibular gland.
我们采用酶消化法和蛋白质印迹分析法，对猴下颌下腺细胞膜和细胞外蛋白聚糖的特性进行了研究。分别从下颌下腺的磷酸盐缓冲盐水（PBS）可溶性部分和微粒体部分提取细胞外蛋白聚糖和细胞膜蛋白聚糖，并通过离子交换色谱法进行部分纯化。醋酸纤维素膜电泳以及用3G10单克隆抗体或6B6单克隆抗体进行免疫印迹分析表明，细胞外蛋白聚糖包含几种硫酸乙酰肝素蛋白聚糖（核心蛋白分子量分别为117、67、47和35 kDa）和一种硫酸皮肤素蛋白聚糖（核心蛋白分子量为45 - 50 kDa），而细胞膜蛋白聚糖仅包含一种核心蛋白分子量为69 kDa的硫酸乙酰肝素蛋白聚糖。这些结果表明，几种细胞外硫酸乙酰肝素蛋白聚糖可能是细胞膜蛋白聚糖从细胞表面释放的胞外域。提示细胞膜和细胞外蛋白聚糖可能在正常下颌下腺中调节生长因子的功能和微环境。
