SOLUTION STRUCTURE OF THE ANTICODON-BINDING DOMAIN OF ESCHERICHIA-COLI LYSYL-TRANSFER-RNA SYNTHETASE AND STUDIES OF ITS INTERACTION WITH TRNA(LYS)
SOLUTION STRUCTURE OF THE ANTICODON-BINDING DOMAIN OF ESCHERICHIA-COLI LYSYL-TRANSFER-RNA SYNTHETASE AND STUDIES OF ITS INTERACTION WITH TRNA(LYS)
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DOI：
10.1006/jmbi.1995.0539
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发表时间：
1995-10-13
影响因子：
5.6
通讯作者：
DARDEL, F
中科院分区：
文献类型：
--
作者：
COMMANS, S;PLATEAU, P;DARDEL, F
对应于大肠杆菌赖氨酰-tRNA合成酶种类的残基31至149的蛋白质结构域被表达并被N-15标记，所述大肠杆菌赖氨酰-tRNA合成酶种类对应于lysS基因。该结构域的 H-1 和 N-15 NMR 共振归属是通过二维和三维同核和异核光谱获得的。使用距离几何和模拟退火，可以使用 701 个 NOE 和 86 个二面角约束来计算三维结构。它由五链反平行β-桶组成，末端有三个α-螺旋。该结构与 lysU 基因表达的其他大肠杆菌赖氨酰-tRNA 合成酶种类的 N 端结构域非常相似，并且与天冬氨酰-tRNA 合成酶相应区域观察到的折叠高度同源。结果表明，分离的赖氨酰-tRNA 合成酶 N 端片段可以与 tRNA(Lys) 以及模拟反密码子序列的聚 (U) 相互作用。鉴定了参与这些相互作用的氨基酸残基，并且就聚U而言，对许多特定的蛋白质-RNA接触进行了表征。 tRNA (Lys) 的特异性识别涉及一组四个结构明确的芳香族残基，锚定在 β 链上，碱性残基位于周围的环上。这种组织让人想起其他 RNA 结合蛋白，例如 U1A 小核核糖核蛋白。 (C) 1995 学术出版社有限公司
A protein domain corresponding to residues 31 to 149 of the E. coli Lysyl-tRNA synthetase species corresponding to the lysS gene was expressed and N-15-labelled. H-1 and N-15 NMR resonance assignments for this domain were obtained by two-dimensional and three-dimensional homonuclear and heteronuclear spectroscopy. Using distance geometry and simulated annealing, a three-dimensional structure could be calculated using 701 NOE and 86 dihedral angle restraints. It is composed of a five-stranded antiparallel beta-barrel capped by three alpha-helices at its ends. This structure closely resembles that of the N-terminal domain of the other E. coli lysyl-tRNA synthetase species expressed from the lysU gene and is highly homologous to the fold observed for the corresponding region of aspartyl-tRNA synthetase. It is shown that the isolated N-terminal fragment of lysyl-tRNA synthetase can interact with tRNA(Lys) as well as with poly (U), which mimics the anticodon sequence. Amino acid residues involved in these interactions were identified and, in the case of poly-U, a number of specific protein-RNA contacts were characterized. Specific recognition of tRNA(Lys) involves a cluster of four structurally well-defined aromatic residues, anchored on the beta-strands, and basic residues located on the surrounding loops. This organization is reminiscent of other RNA binding proteins, such as the U1A small nuclear ribonucleoprotein. (C) 1995 Academic Press Limited