Detection of dissolved protein molecules in oceanic waters
Detection of dissolved protein molecules in oceanic waters
复制标题
检测海水中溶解的蛋白质分子
DOI：
10.1016/0304-4203(95)00061-5
复制
发表时间：
1995
期刊：
影响因子：
3
通讯作者：
E. Tanoue
中科院分区：
文献类型：
--
作者：
E. Tanoue
描述了一种提取和检测海洋沃茨中溶解蛋白质分子的方法。该方法包括三个独立的步骤：（1）用切向流超滤法从海水中粗浓缩溶解蛋白，（2）用三氯乙酸沉淀法进一步浓缩和纯化溶解蛋白，（3）用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法分离和检测溶解蛋白。溶解的蛋白质被成功地提取与这种技术，相对有限的蛋白质分子（少于30）被分离和可视化的凝胶上的主要带在分析水从站位于亚北极到热带太平洋。大多数蛋白质的分子量范围为14至66千道尔顿（kDa）。海水中溶解蛋白质的电泳图谱在水平和垂直方向上都有变化，但在所有样品中均发现了一些蛋白质分子。检测到的主要蛋白质是相对纯的，并且以高水平存在。相对有限数量的蛋白质的大量积累导致了这样的假设：构成海水中溶解蛋白质成分大部分的特定蛋白质来自特定来源，并对海洋有机氮库做出了定量贡献。
A method for the extraction and detection of dissolved protein molecules in oceanic waters is described. The procedure involves three separate steps: (1) crude concentration of dissolved protein from seawater by tangential flow ultrafiltration, (2) further concentration and purification of dissolved protein by precipitation with trichloroacetic acid, and (3) separation and detection of dissolved proteins by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Dissolved proteins were successfully extracted with this technique; a relatively limited number of protein molecules (less than 30) were separated and visualized on gels as major bands during the analysis of water from stations located from the subarctic to the tropical Pacific. The majority of proteins had molecular masses ranging from 14 to 66 kilodaltons (kDa). Electrophoretic patterns of dissolved proteins changed both horizontally and vertically in seawater, but some protein molecules were found in all the samples examined. The major proteins detected were relatively pure and were present at high levels. The accumulation of appreciable amounts of a relatively limited number of proteins leads to the hypothesis that particular proteins that make up the majority of the dissolved protein components in seawater are derived from specific sources and contribute quantitatively to the oceanic pool of organic nitrogen.