Probing RegA/RNA interactions using electrospray ionization-fourier transform ion cyclotron resonance-mass spectrometry.
Probing RegA/RNA interactions using electrospray ionization-fourier transform ion cyclotron resonance-mass spectrometry.
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使用电喷雾电离-傅里叶变换离子回旋共振-质谱法探测 RegA/RNA 相互作用。
DOI：
10.1006/abio.1998.2753
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发表时间：
1998
影响因子：
2.9
通讯作者：
Sinha,N
中科院分区：
文献类型：
--
作者：
Liu,C;Tolić,LP;Hofstadler,SA;Harms,AC;Smith,RD;Kang,C;Sinha,N
利用电喷雾电离-傅立叶变换离子回旋共振-质谱研究了噬菌体T4 regA蛋白（一种独特的翻译调节因子）与不同大小和序列的RNA的相互作用。使用非常温和的界面条件，regA/RNA复合物与1：1的结合化学计量观察到所有四个目标RNA的研究，与溶液结合研究。靶RNA及其降解产物与regA的竞争性结合表明，在某些情况下，单个核苷酸的丢失导致结合亲和力的急剧变化。regA与四种靶RNA的竞争性结合揭示了与先前的体外抑制实验所建议的相似的相对结合亲和力顺序。使用持续的非共振辐射碰撞诱导解离的regA/RNA复合物建议直接获得有关regA结合域的信息的潜力。
The interactions of bacteriophage T4 regA protein, a unique translational regulator, with RNAs of various size and sequence were studied using electrospray ionization–Fourier transform ion cyclotron resonance–mass spectrometry. Using very gentle interface conditions, regA/RNA complexes with a 1:1 binding stoichiometry were observed for all four target RNAs studied, consistent with solution binding studies. Competitive binding of target RNAs and their degradation products with regA demonstrated that the loss of a single nucleotide resulted in a dramatic change in binding affinity in some cases. Competitive binding of regA with four target RNAs revealed similar relative binding affinity order to that suggested by previousin vitrorepression experiments. The use of sustained off-resonance irradiation for collisionally induced dissociation of a regA/RNA complex suggested the potential for directly obtaining information regarding the regA binding domain.