解决核磁共振结构和动力学的主要支柱冷休克蛋白(CspA)从大肠杆菌:证据单链rna结合构象动态的网站。
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冯W,特R,齐默尔曼,Inouye M, Montelione GT
解决核磁共振结构和动力学的主要支柱冷休克蛋白(CspA)从大肠杆菌:证据单链rna结合构象动态的网站。
生物化学。1998年8月4日,37 (31):10881 - 96。doi: 10.1021 / bi980269j。
- PubMed ID
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9692981 (在PubMed]
- 文摘
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从大肠杆菌主要冷休克蛋白(CspA)是一种单链核酸结合蛋白在冷应激反应产生。我们曾报道其整体链折叠由核磁共振光谱学(纽克K。冯,W。江,W。特,R。爱默生,美国D。Inouye, M。,Montelione g . t . (1994) Proc。国家的。学会科学。美国91年,5114 - 5118]。这里描述的完整分析1 h、13 c和15 n CspA共振作业,一起细化解决方案基于699构象限制核磁共振结构和分析基于15 n的骨干动力学弛豫率测量。一组广泛的triple-resonance NMR实验获取的骨干和侧链共振作业统一进行13 c - 15 n-enriched CspA。 Using a subset of these triple-resonance experiments, the computer program AUTOASSIGN provided automatic analysis of sequence-specific backbone N, Calpha, C', HN, Halpha, and side chain Cbeta resonance assignments. The remaining 1H, 13C, and 15N resonance assignments for CspA were then obtained by manual analysis of additional NMR spectra. Dihedral angle constraints and stereospecific methylene Hbeta resonance assignments were determined using a new conformational grid search program, HYPER, and used together with longer-range constraints as input for three-dimensional structure calculations. The resulting solution NMR structure of CspA is a well-defined five-stranded beta-barrel with surface-exposed aromatic groups that form a single-stranded nucleic acid-binding site. Backbone dynamics of CspA have also been characterized by 15N T1, T2, and heteronuclear 15N-1H NOE measurements and analyzed using the extended Lipari-Szabo formalism. These dynamic measurements indicate a molecular rotational correlation time taum of 4.88 +/- 0.04 ns and provide evidence for fast time scale (taue < 500 ps) dynamics in surface loops and motions on the microsecond to millisecond time scale within the proposed nucleic acid-binding epitope.