Horwitz A L, Dorfman A
J Cell Biol. 1968 Aug;38(2):358-68. doi: 10.1083/jcb.38.2.358.
Microsomes from embryonic cartilage have been subfractionated to yield smooth microsomes and rough microsomes. The in vitro enzymic activities involved in chondroitin sulfate biosynthesis have been assayed in these subfractions. The results demonstrate that all of the activities necessary for linkage to protein as well as for completion of the polysaccharide chain are present in both the rough and smooth fractions. Only in the case of the polymerization of N-acetylgalactosamine and glucuronic acid could enzyme assays be done independent of endogenous acceptor. This enzyme(s) was equally distributed between the rough and smooth fractions. The activities for the addition of xylose and galactose to protein were highest in the rough fraction while that for sulfation was highest in the smooth fraction. These findings suggest that polysaccharide chain-initiation occurs in the rough endoplasmic reticulum and that chain completion occurs in the smooth reticulum. This pattern is consistent with modern theories of synthesis, transfer, and export of extracellular macromolecules.
来自胚胎软骨的微粒体已被分级分离,以产生光滑微粒体和粗糙微粒体。已在这些亚组分中测定了硫酸软骨素生物合成中涉及的体外酶活性。结果表明,与蛋白质连接以及多糖链完成所需的所有活性在粗糙和光滑组分中均存在。只有在N-乙酰半乳糖胺和葡萄糖醛酸聚合的情况下,酶测定才能独立于内源性受体进行。这种酶在粗糙和光滑组分中分布均匀。将木糖和半乳糖添加到蛋白质中的活性在粗糙组分中最高,而硫酸化活性在光滑组分中最高。这些发现表明多糖链起始发生在粗糙内质网中,而链的完成发生在光滑内质网中。这种模式与细胞外大分子合成、转运和输出的现代理论一致。