Biotin-streptavidin-labeled oligonucleotides as probes of helicase mechanisms
Biotin-streptavidin-labeled oligonucleotides as probes of helicase mechanisms
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DOI：
10.1006/meth.2000.1116
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发表时间：
2001-02-01
期刊：
影响因子：
4.8
通讯作者：
Raney, KD
中科院分区：
文献类型：
--
作者：
Morris, PD;Tackett, AJ;Raney, KD
解旋酶使用来自ATP水解的能量通过解旋双链核酸来催化单链核酸的形成。ATP依赖性反应可以被分解为至少两个步骤：双链体的解链和酶沿着核酸晶格的移位。每个步骤都给研究带来困难，因为反应的明确终点并不总是可用的。例如，易位涉及酶从一个点沿着晶格移动到一个新的位置，而核酸的化学结构没有净变化。因此，已经开发了新的测定法，其中核酸被修饰为含有阻碍酶移位的"蛋白质块"。为了制备这样的蛋白质块，已经使用生物素-链霉亲和素，因为生物素可以通过化学合成容易地掺入核酸中。几个应用生物素-链霉亲和素标记的寡核苷酸的解旋酶机制的研究。(C)北京：科学出版社.
Helicases use the energy from ATP hydrolysis to catalyze formation of single-stranded nucleic acids by unwinding double-stranded nucleic acids. The ATP-dependent reaction can be broken down into at least two steps: melting of the duplex and translocation of the enzyme along the nucleic acid lattice. Each step presents difficulties for study because clear endpoints for the reactions are not always available. For example, translocation involves the movement of the enzyme from one point along the lattice to a new position, with no net change in chemical structure of the nucleic acid. Hence, new assays have been developed in which the nucleic acid is modified to contain a "protein block" that impedes translocation of the enzyme. To prepare such protein blocks, biotin-streptavidin has been used due to the ease with which the biotin can be incorporated into nucleic acids by chemical synthesis. Several applications of oligonucleotides labeled with biotin-streptavidin for the study of helicase mechanisms are described. (C) 2001 Academic Press.