Nature子刊：目睹细胞中的折叠
研究人员在11月在线出版的《自然—化学生物学》期刊上报告说，与试管中的研究相比，一种亲眼目睹蛋白质在细胞中折叠的方法也许能更好地帮助我们深入认识蛋白质的不同功能。
蛋白质是由线性的高分子链经过折叠成三维结构后形成的。蛋白质的功能取决于结构，因此了解这一折叠过程非常重要。尽管这种折叠过程在人工实验环境下已被广泛研究，但细胞的极复杂的真实环境却会影响蛋白质的三维结构。
Schepartz和同事将一种接合已知荧光分子的氨基酸标签打破成两段，让它们分别粘贴在分离的蛋白质链的两端。当蛋白质成功折叠时，这些片段就组合在一起，发出荧光；而当蛋白质不能正常折叠时，荧光信号就不会出现。（科学时报）
原始出处:
Nature Chemical Biology
Published online: 4 November 2007 | doi:10.1038/nchembio.2007.49
Surveying polypeptide and protein domain conformation and association with FlAsH and ReAsH
Nathan W Luedtke1,3, Rachel J Dexter1, Daniel B Fried1 & Alanna Schepartz1,2
Recombinant polypeptides and protein domains containing two cysteine pairs located distal in primary sequence but proximal in the native folded or assembled state are labeled selectively in vitro and in mammalian cells using the profluorescent biarsenical reagents FlAsH-EDT2 and ReAsH-EDT2. This strategy, termed bipartite tetracysteine display, enables the detection of protein-protein interactions and alternative protein conformations in live cells. As proof of principle, we show that the equilibrium stability and fluorescence intensity of polypeptide–biarsenical complexes correlates with the thermodynamic stability of the protein fold or assembly. Destabilized protein variants form less stable and less bright biarsenical complexes, which allows discrimination of live cells expressing folded polypeptide and protein domains from those containing disruptive point mutations. Bipartite tetracysteine display may provide a means to detect early protein misfolding events associated with Alzheimer's disease, Parkinson's disease and cystic fibrosis; it may also enable high-throughput screening of compounds that stabilize discrete protein folds.
- Department of Chemistry, Yale University, 225 Prospect Street, New Haven, Connecticut 06520-8107, USA.
- Department of Chemistry and Molecular, Cellular, and Developmental Biology, Yale University, 219 Prospect Street, New Haven, Connecticut 06520-8107, USA.
- Present address: Universität Zürich, Organisch-chemisches Institut, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
Correspondence to: Alanna Schepartz1,2 Email: alanna.schepartz@yale.edu
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