Keller R K, Swank G D
Biochim Biophys Acta. 1981 Feb 18;673(1):93-100.
In order to determine if the rates of intracellular transport of oviduct secretory proteins are coordinate, oviduct minces were first incubated with either [3H]methionine or [3H]leucine. 2 h later, the minces were exposed to either [35S]methionine or [14C]leucine in addition to the 3H-labeled amino acid. After incubating for an additional 90-120 min, aliquots of the media were subjected to immunoprecipitation to isolate ovalbumin, conalbumin and lysozyme. From the 3H/35S or 3H/14C ratios in the isolated proteins, relative rates of intracellular transport were determined. The data obtained indicate that conalbumin and lysozyme exhibit similar rates of intracellular transport, whereas ovalbumin lags behind these proteins considerably. The basis for this effect is discussed in light of current knowledge concerning the synthesis and secretion of ovalbumin.
为了确定输卵管分泌蛋白的细胞内运输速率是否协调,首先将输卵管切碎组织与[3H]甲硫氨酸或[3H]亮氨酸一起孵育。2小时后,除了3H标记的氨基酸外,将切碎组织暴露于[35S]甲硫氨酸或[14C]亮氨酸中。再孵育90 - 120分钟后,取部分培养基进行免疫沉淀以分离卵清蛋白、伴清蛋白和溶菌酶。根据分离出的蛋白质中的3H/35S或3H/14C比率,确定细胞内运输的相对速率。所获得的数据表明,伴清蛋白和溶菌酶表现出相似的细胞内运输速率,而卵清蛋白在这些蛋白质之后有相当大的滞后。根据目前关于卵清蛋白合成和分泌的知识讨论了这种效应的基础。