Emtage J S, Lawson D E, Kodicek E
Biochem J. 1974 May;140(2):239-47. doi: 10.1042/bj1400239.
Undegraded polyribosome preparations may be obtained from chick intestinal mucosa if ribonuclease activity is strictly controlled. This is best achieved by homogenization of the mucosa directly in rat liver cell-sap. 2. The extent of amino acid incorporation by chick intestinal polyribosomes is greatly influenced by the source of the cell-sap. Sephadex-treated intestinal cell-sap caused impaired incorporation and release of completed polypeptide chains, whereas Sephadex-treated rat liver cell-sap promoted the polymerization of up to 90 amino acids per ribosome. Under optimum conditions 30-35% of the nascent polypeptide chains are completed and released. 3. The preparation of an antiserum against the calcium-binding protein formed in response to vitamin D is described. It is shown that the antiserum is highly specific for calcium-binding protein. 4. This antiserum was used to investigate the ability of chick intestinal polyribosomes to synthesize calciumbinding protein. Only polyribosomes from chicks receiving vitamin D have the ability to synthesize calcium-binding protein. Moreover, the product formed in vitro has the same electrophoretic mobility as calcium-binding protein synthesized in vivo. 5. It is concluded that one of the main functions of vitamin D in the small intestine is to induce the synthesis de novo of calcium-binding protein.
如果严格控制核糖核酸酶的活性,就可以从鸡的肠黏膜中获得未降解的多核糖体制剂。最好的方法是将黏膜直接在大鼠肝细胞液中匀浆。2. 鸡肠多核糖体掺入氨基酸的程度受细胞液来源的影响很大。经葡聚糖凝胶处理的肠细胞液会导致已完成的多肽链掺入和释放受损,而经葡聚糖凝胶处理的大鼠肝细胞液则促进每个核糖体聚合多达90个氨基酸。在最佳条件下,30%至35%的新生多肽链会完成并释放。3. 描述了一种针对维生素D诱导形成的钙结合蛋白的抗血清的制备方法。结果表明,该抗血清对钙结合蛋白具有高度特异性。4. 该抗血清用于研究鸡肠多核糖体合成钙结合蛋白的能力。只有接受维生素D的鸡的多核糖体才有合成钙结合蛋白的能力。此外,体外形成的产物与体内合成的钙结合蛋白具有相同的电泳迁移率。5. 得出的结论是,维生素D在小肠中的主要功能之一是诱导从头合成钙结合蛋白。