Mobility of Histidine Side Chains Analyzed with 15N NMR Relaxation and Cross-Correlation Data: Insight into Zinc-Finger-DNA Interactions.

Mobility of Histidine Side Chains Analyzed with 15N NMR Relaxation and Cross-Correlation Data: Insight into Zinc-Finger-DNA Interactions.
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使用 15N NMR 弛豫和互相关数据分析组氨酸侧链的迁移率:深入了解锌指-DNA 相互作用。

DOI:
10.1021/acs.jpcb.9b03132
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发表时间:
2019
期刊:
The journal of physical chemistry. B
影响因子:
--
通讯作者:
Iwahara,Junji
Iwahara,Junji
中科院分区:
--
文献类型:
--
作者:
Kemme,CatherineA;Luu,RossH;Chen,Chuanying;Pletka,ChanningC;Pettitt,BMontgomery;Iwahara,Junji

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由于化学交换,蛋白质组氨酸(His)侧链的迁移性通常难以通过核磁共振光谱分析。使用不受化学交换影响的核磁共振方法,我们研究了Egr-1锌指- DNA复合物中直接与DNA磷酸盐相互作用的His咪唑NH基团的内部运动。在此方法中,我们分析了15n化学位移各向异性和15n - 1h偶极子-偶极子弛豫干涉的横向和纵向互相关率,以及15n纵向弛豫率和异核Overhauser效应在两种磁场强度下的数据。我们发现直接与DNA磷酸盐相互作用的锌配位His侧链在移动性上受到强烈限制。这与精氨酸和赖氨酸侧链形成对比,尽管它们与相同复合物中的DNA磷酸盐相互作用,但仍保持高迁移率。讨论了侧链迁移率对分子缔合的熵效应。
Due to chemical exchange, the mobility of histidine (His) side chains of proteins is typically difficult to analyze by NMR spectroscopy. Using an NMR approach that is uninfluenced by chemical exchange, we investigated internal motions of the His imidazole NH groups that directly interact with DNA phosphates in the Egr-1 zinc-finger–DNA complex. In this approach, the transverse and longitudinal cross-correlation rates for15N chemical shift anisotropy and15N–1H dipole–dipole relaxation interference were analyzed together with15N longitudinal relaxation rates and heteronuclear Overhauser effect data at two magnetic field strengths. We found that the zinc-coordinating His side chains directly interacting with DNA phosphates are strongly restricted in mobility. This makes a contrast to the arginine and lysine side chains that retain high mobility despite their interactions with DNA phosphates in the same complex. The entropic effects of side-chain mobility on the molecular association are discussed.
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