Luminescence resonance energy transfer analysis of RNA polymerase complexes
Luminescence resonance energy transfer analysis of RNA polymerase complexes
复制标题
DOI：
10.1006/meth.2001.1214
复制
发表时间：
2001-09-01
期刊：
影响因子：
4.8
通讯作者：
Heyduk, T
中科院分区：
文献类型：
--
作者：
Heyduk, T
共振能量转移允许在各种溶液条件下测量原子尺度的距离。发光共振能量转移(LRET)是一种以稀土络合物为探针的能量转移测量方法。镧系元素发射的不同寻常的性质，特别是其长的微秒级寿命，为利用生物样品测量能量转移提供了几个优点。LRET的独特功能之一是能够在标记大分子存在严重异质性的条件下测量能量转移。LRET的这一特点是本文特别强调的。我们在这里描述了LRET方法，特别注意使用敏化的受主发射来确定能量转移的效率。虽然我们在鉴定大肠杆菌RNA聚合酶复合体时使用了这项技术，但它很容易与基本上任何蛋白质的研究兼容。(C)2001年学术出版社。
Resonance energy transfer allows measurement of atomic-scale distances under a variety of solution conditions. Luminescence resonance energy transfer (LRET) is a variant of energy transfer measurement in which lanthanide chelates are used as the probes. The unusual properties of lanthanide emission, in particular their long microsecond-scale lifetimes, offer several advantages for energy transfer measurements with biological samples. One of the unique features of LRET is the ability to measure energy transfer under conditions where severe heterogeneity of labeled macromolecules exists. This feature of LRET is the special emphasis of this article. We describe here LRET methodology with a particular attention to using sensitized acceptor emission to determine efficiency of energy transfer. Although we employed this technique in the characterization of Escherichia coli RNA polymerase complexes it Is readily compatible with the study of essentially any protein. (C) 2001 Academic Press.