Site-directed spin labeling measurements of nanometer distances in nucleic acids using a sequence-independent nitroxide probe.
Site-directed spin labeling measurements of nanometer distances in nucleic acids using a sequence-independent nitroxide probe.
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DOI：
10.1093/nar/gkl546
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发表时间：
2006
影响因子：
14.9
通讯作者：
Qin PZ
中科院分区：
文献类型：
--
作者：
Cai Q;Kusnetzow AK;Hubbell WL;Haworth IS;Gacho GP;Van Eps N;Hideg K;Chambers EJ;Qin PZ
在定点自旋标记(SDSL)中，局部结构和动态信息是通过固定在大分子上的稳定氮氧自由基的电子顺磁共振(EPR)谱获得的。对氮氧化物对之间的电子自旋偶极相互作用的分析得到了氮氧化物之间的距离，这提供了定量的结构信息。脉冲电子顺磁共振方法的发展使这种生物分子的距离测量达到了70 Å，从而为立体定向激光全球结构图谱的绘制提供了可能。这项研究评估了使用氮氧化物(指定为R5)的SDSL距离测量，该氮氧化物可以以有效和经济的方式连接到任意DNA或RNA序列上的硫代磷酸盐主干位置。将R5对连接到具有已知核磁共振结构的十二聚体双链的选定位置，并使用双电子-电子共振(DER)测量了从20到40 Å的8个距离。测量的距离与根据对核磁共振结构中空间上允许的R5构象的搜索计算出的预测值具有很强的相关性(R2=0.98)，从而证明了使用R5进行准确的距离测量。此外，还对42kD的DNA进行了距离测量。这些结果使R5成为DNA和DNA-蛋白质复合体的全球结构图谱的独立于序列的探针。
In site-directed spin labeling (SDSL), local structural and dynamic information is obtained via electron paramagnetic resonance (EPR) spectroscopy of a stable nitroxide radical attached site-specifically to a macromolecule. Analysis of electron spin dipolar interactions between pairs of nitroxides yields the inter-nitroxide distance, which provides quantitative structural information. The development of pulse EPR methods has enabled such distance measurements up to 70 Å in bio-molecules, thus opening up the possibility of SDSL global structural mapping. This study evaluates SDSL distance measurement using a nitroxide (designated as R5) that can be attached, in an efficient and cost-effective manner, to a phosphorothioate backbone position at arbitrary DNA or RNA sequences. R5 pairs were attached to selected positions of a dodecamer DNA duplex with a known NMR structure, and eight distances, ranging from 20 to 40 Å, were measured using double electron-electron resonance (DEER). The measured distances correlated strongly (R2 = 0.98) with the predicted values calculated based on a search of sterically allowable R5 conformations in the NMR structure, thus demonstrating accurate distance measurements using R5. Furthermore, distance measurement in a 42 kD DNA was demonstrated. The results establish R5 as a sequence-independent probe for global structural mapping of DNA and DNA–protein complexes.
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影响因子：
5.3
作者：
DEB, S;DELUCIA, AL;TEGTMEYER, P
通讯作者：
TEGTMEYER, P
影响因子：
15
作者：
Borbat, PP;Mchaourab, HS;Freed, JH
通讯作者：
Freed, JH
影响因子：
2.2
作者：
Chiang, YW;Borbat, PP;Freed, JH
通讯作者：
Freed, JH
影响因子：
0.8
作者：
Chen, SY;Nie, JJ;Chen, YZ
通讯作者：
Chen, YZ
DOI：
10.1107/s056774088200510x
发表时间：
1982-01-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION B-STRUCTURAL SCIENCE
影响因子：
--
作者：
GOLDSTEIN, BM
通讯作者：
GOLDSTEIN, BM