Bartlett S F, Lagercrantz H, Smith A D
Department of Pharmacology, University of Oxford, Oxford OX1 3QT, England.
Neuroscience. 1976 Aug;1(4):339-44. doi: 10.1016/0306-4522(76)90061-0.
Polyacrylamide gel electrophoresis of a soluble extract of purified large dense-cored vesicles (noradrenergic vesicles) from ox splenic nerve revealed 13 proteins, 7 of which had mobilities close to those of seven of the soluble proteins in adrenal chromaffin granules. Two of these proteins had the same mobilities as dopamine beta-hydroxylase and chromogranin A, respectively. The relative staining intensities of the latter two proteins were different: in noradrenergic vesicles, there was more dopamine beta-hydroxylase than chromogranin A; whereas in chromaffin granules, chromogranin A was the major protein. Gel electrophoresis of the water-insoluble proteins of noradrenergic vesicles, dissolved in sodium dodecylsulphate, revealed 5 proteins, 5 of which had mobilities close to those of 5 proteins present in the membranes of chromaffin granules. Three of these proteins had mobilities similar to those of dopamine beta-hydroxylase, chromogranin A and chromomembrin B, respectively. One of the proteins was probably serum albumin, which was also present as a contaminant in the soluble extract of noradrenergic vesicles. These findings are consistent with earlier studies in which dopamine beta-hydroxylase, chromogranin A and chromomembrin B have been identified as constituents of noradrenergic vesicles by enzymatic or immunochemical assay methods. They also indicate further qualitative similarities between the noradrenergic vesicle and the chromaffin granule, in that a total of 7 soluble and 5 insoluble proteins might be common to both particles. However, gel electrophoresis also confirms that quantitative differences exist between the relative proportions of the soluble proteins and shows that dopamine beta-hydroxylase is the major soluble protein of the noradrenergic vesicles isolated from splenic nerve trunks.
对从牛脾神经中纯化得到的大致密核心囊泡（去甲肾上腺素能囊泡）的可溶性提取物进行聚丙烯酰胺凝胶电泳，结果显示有13种蛋白质，其中7种蛋白质的迁移率与肾上腺嗜铬颗粒中7种可溶性蛋白质的迁移率相近。其中两种蛋白质的迁移率分别与多巴胺β-羟化酶和嗜铬粒蛋白A相同。后两种蛋白质的相对染色强度不同：在去甲肾上腺素能囊泡中，多巴胺β-羟化酶比嗜铬粒蛋白A多；而在嗜铬颗粒中，嗜铬粒蛋白A是主要蛋白质。对溶解于十二烷基硫酸钠中的去甲肾上腺素能囊泡的水不溶性蛋白质进行凝胶电泳，结果显示有5种蛋白质，其中5种蛋白质的迁移率与嗜铬颗粒膜中存在的5种蛋白质的迁移率相近。其中3种蛋白质的迁移率分别与多巴胺β-羟化酶、嗜铬粒蛋白A和嗜铬膜蛋白B的迁移率相似。其中一种蛋白质可能是血清白蛋白，它在去甲肾上腺素能囊泡的可溶性提取物中也作为污染物存在。这些发现与早期的研究一致，在早期研究中，多巴胺β-羟化酶、嗜铬粒蛋白A和嗜铬膜蛋白B已通过酶促或免疫化学分析方法被鉴定为去甲肾上腺素能囊泡的成分。它们还进一步表明去甲肾上腺素能囊泡和嗜铬颗粒在质量上存在相似性，因为这两种颗粒可能共有7种可溶性蛋白质和5种不溶性蛋白质。然而，凝胶电泳也证实了可溶性蛋白质的相对比例存在定量差异，并表明多巴胺β-羟化酶是从脾神经干分离得到的去甲肾上腺素能囊泡的主要可溶性蛋白质。
