Whitaker M
Department of Physiological Sciences, Medical School, Framlington Place, Newcastle upon Tyne, NE2 4HH UK.
Bioessays. 2000 Feb;22(2):180-7. doi: 10.1002/(SICI)1521-1878(200002)22:2<180::AID-BIES9>3.0.CO;2-M.
A cell's biochemistry is now known to be the biochemistry of molecular machines, that is, protein complexes that are assembled and dismantled in particular locations within the cell as needed. One important element in our understanding has been the ability to begin to see where proteins are in cells and what they are doing as they go about their business. Accordingly, there is now a strong impetus to discover new ways of looking at the workings of proteins in living cells. Although the use of fluorescent tags to track individual proteins in cells has a long history, the availability of laser-based confocal microscopes and the imaginative exploitation of the green fluorescent protein from jellyfish have provided new tools of great diversity and utility. It is now possible to watch a protein bind its substrate or its partners in real time and with submicron resolution within a single cell. The importance of processes of self-organisation represented by protein folding on the one hand and subcellular organelles on the other are well recognised. Self-organisation at the intermediate level of multimeric protein complexes is now open to inspection. BioEssays 22:180-187, 2000.
如今已知细胞的生物化学就是分子机器的生物化学,也就是说,蛋白质复合物会根据需要在细胞内的特定位置组装和拆解。我们理解过程中的一个重要因素是能够开始观察蛋白质在细胞中的位置以及它们在执行任务时的行为。因此,现在有一股强大的动力去发现观察活细胞中蛋白质作用方式的新方法。尽管使用荧光标签追踪细胞中的单个蛋白质已有很长历史,但基于激光的共聚焦显微镜的出现以及对水母绿色荧光蛋白的巧妙利用提供了种类繁多且实用的新工具。现在有可能在单个细胞内以亚微米分辨率实时观察一种蛋白质与其底物或伙伴的结合。一方面由蛋白质折叠、另一方面由亚细胞器所代表的自组织过程的重要性已得到充分认识。多聚体蛋白质复合物中间层面的自组织现在也可供研究。《生物论文》22:180 - 187,2000年。