Shub M D, Pang K Y, Swann D A, Walker W A
Biochem J. 1983 Nov 1;215(2):405-11. doi: 10.1042/bj2150405.
Mucus glycoproteins from newborn and adult rat small intestine were radiolabelled in vivo with Na2 35SO4 and isolated from mucosal homogenates by using Sepharose 4B column chromatography followed by CsCl-density-gradient centrifugation. Non-covalently bound proteins, lipids and nucleic acids were not detected in the purified glycoproteins. Amino acid, carbohydrate and sulphate compositions were similar to chemical compositions reported for other intestinal mucus glycoproteins, as were sedimentation properties. There were, however, important differences in the chemical and physical characteristics of the mucus glycoproteins from newborn and adult animals. The buoyant density in CsCl was higher for the glycoproteins from newborn rats (1.55 g/ml versus 1.47 g/ml). On sodium dodecyl sulphate/polyacrylamide/agarose-gel electrophoresis, the glycoprotein from newborn rats had a greater mobility than the adult-rat sample. Although both preparations had similar general amino acid compositions, variations were observed for individual amino acids. The total protein content was greater in the glycoprotein from newborn animals (27%, w/w, versus 18%, w/w). The molar ratio of carbohydrate to protein was less in the newborn, primarily owing to a decreased fucose and N-acetylgalactosamine content. Comparison of the molar ratio of fucose and sialic acid to galactose for both glycoproteins demonstrated a reciprocal relationship similar to that described by Dische [(1963) Ann. N.Y. Acad. Sci. 106, 259-270]. The sulphate content was greater in the glycoprotein from newborn rats (5.5%, w/w, versus 0.9%, w/w). Both had similar sedimentation coefficients in a dissociative solvent. These results suggest an age-related difference in the types of mucus glycoproteins synthesized by small intestine.
新生大鼠和成年大鼠小肠的黏液糖蛋白在体内用Na2 35SO4进行放射性标记,然后通过琼脂糖4B柱色谱法,接着用CsCl密度梯度离心法从黏膜匀浆中分离出来。在纯化的糖蛋白中未检测到非共价结合的蛋白质、脂质和核酸。氨基酸、碳水化合物和硫酸盐组成与其他肠道黏液糖蛋白报道的化学组成相似,沉降特性也相似。然而,新生动物和成年动物的黏液糖蛋白在化学和物理特性上存在重要差异。新生大鼠糖蛋白在CsCl中的浮力密度更高(1.55 g/ml对1.47 g/ml)。在十二烷基硫酸钠/聚丙烯酰胺/琼脂糖凝胶电泳中,新生大鼠的糖蛋白比成年大鼠样品具有更大的迁移率。尽管两种制剂的总体氨基酸组成相似,但个别氨基酸存在差异。新生动物糖蛋白中的总蛋白含量更高(27%,w/w,对18%,w/w)。新生动物中碳水化合物与蛋白质的摩尔比更低,主要是由于岩藻糖和N-乙酰半乳糖胺含量降低。两种糖蛋白中岩藻糖和唾液酸与半乳糖的摩尔比比较显示出与迪谢所描述的相似的反比关系[(1963年)《纽约科学院学报》106,259 - 270]。新生大鼠糖蛋白中的硫酸盐含量更高(5.5%,w/w,对0.9%,w/w)。在解离溶剂中两者具有相似的沉降系数。这些结果表明小肠合成的黏液糖蛋白类型存在与年龄相关的差异。