McQuillan D J, Findlay D M, Hocking A M, Yanagishita M, Midura R J, Hascall V C
Department of Paediatrics, University of Melbourne, Royal Children's Hospital, Parkville, Victoria, Australia.
Biochem J. 1991 Jul 1;277 ( Pt 1)(Pt 1):199-206. doi: 10.1042/bj2770199.
The proteoglycans synthesized by an osteoblast-like cell line of rat origin (UMR 106-01) were defined after biosynthetic labelling with [35S]sulphate and [3H]glucosamine. Newly synthesized labelled proteoglycans were characterized by differential enzymic digestion in combination with analytical gel filtration and SDS/PAGE. UMR 106-01 cells were found to synthesize three major species of proteoglycan: a large chondroitin sulphate proteoglycan of Mr approximately 1 x 10(6), with a core protein of Mr approximately 350,000-400,000; a small chondroitin sulphate-containing species of Mr approximately 120,000 with a core protein of Mr 43,000; and a heparan sulphate proteoglycan of Mr approximately 150,000, with a core protein of Mr approximately 80,000. Over 70% of the newly synthesized intact proteoglycan species are associated with the cell layer of near-confluent cells; however, accessibility to trypsin digestion suggests an extracellular location. Chemical characteristics of the proteoglycans and preliminary mRNA hybridization indicate that the small chondroitin sulphate proteoglycan is probably PG II (decorin). The large chondroitin sulphate proteoglycan is most likely related to a hyaluronate-aggregating species from fibroblasts (versican), and the heparan sulphate proteoglycan bears striking similarities to cell-membrane-intercalated species described for a number of cell types.
用[35S]硫酸盐和[3H]葡糖胺进行生物合成标记后,对源自大鼠的成骨细胞样细胞系(UMR 106 - 01)合成的蛋白聚糖进行了鉴定。通过结合分析性凝胶过滤和SDS/PAGE的差异酶消化来表征新合成的标记蛋白聚糖。发现UMR 106 - 01细胞合成三种主要的蛋白聚糖:一种Mr约为1×10(6)的大硫酸软骨素蛋白聚糖,其核心蛋白Mr约为350,000 - 400,000;一种Mr约为120,000的含硫酸软骨素的小蛋白聚糖,其核心蛋白Mr为43,000;以及一种Mr约为150,000的硫酸乙酰肝素蛋白聚糖,其核心蛋白Mr约为80,000。超过70%新合成的完整蛋白聚糖种类与接近汇合细胞的细胞层相关;然而,胰蛋白酶消化的可及性表明其位于细胞外。蛋白聚糖的化学特性和初步的mRNA杂交表明,小硫酸软骨素蛋白聚糖可能是PG II(核心蛋白聚糖)。大硫酸软骨素蛋白聚糖很可能与成纤维细胞的一种透明质酸聚集种类(多功能蛋白聚糖)有关,并且硫酸乙酰肝素蛋白聚糖与多种细胞类型所描述的插入细胞膜的种类有显著相似性。