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A Non-Invasive NMR Method Based on Histidine Imidazoles to Analyze the pH-Modulation of Protein-Nucleic Acid Interfaces

机译:基于组氨酸咪唑的非侵入式NMR方法分析蛋白质-核酸界面的pH调节

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摘要

A useful (2)J(N-H) coupling-based NMR spectroscopic approach is proposed to unveil, at the molecular level, the contribution of the imidazole groups of histidines from RNA/DNA-binding proteins on the modulation of binding to nucleic acids by pH. Such protonation/deprotonation events have been monitored on the single His96 located at the second RNA/DNA recognition motif (RRM2) of T-cell intracellular antigen-1 (TIA-1) protein. The pK(a) values of the His96 ionizable groups were substantially higher in the complexes with short U-rich RNA and T-rich DNA oligonucleotides than those of the isolated TIA-1 RRM2. Herein, the methodology applied to determine changes in pK(a) of histidine side chains upon DNA/RNA binding, gives valuable information to understand the pH effect on multidomain DNA/RNA-binding proteins that shuttle among different cellular compartments.
机译:提出了一种有用的基于(2)J(NH)偶联的NMR光谱方法,以在分子水平上揭示RNA / DNA结合蛋白中组氨酸的咪唑基团对pH调节核酸结合的贡献。此类质子化/去质子化事件已在位于T细胞细胞内抗原1(TIA-1)蛋白第二个RNA / DNA识别基序(RRM2)的单个His96上进行了监测。在具有短的富含U的RNA和富含T的DNA寡核苷酸的复合物中,His96可电离基团的pK(a)值明显高于分离的TIA-1 RRM2。在本文中,用于确定DNA / RNA结合后组氨酸侧链pK(a)变化的方法学提供了有价值的信息,以了解pH对在不同细胞区室间穿梭的多域DNA / RNA结合蛋白的影响。

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