Pyrrolysyl-tRNA synthetase-tRNA(Pyl) structure reveals the molecular basis of orthogonality.
Pyrrolysyl-tRNA synthetase-tRNA(Pyl) structure reveals the molecular basis of orthogonality.
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DOI：
10.1038/nature07611
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发表时间：
2009-02-26
期刊：
影响因子：
64.8
通讯作者：
Nureki, Osamu
中科院分区：
文献类型：
--
作者：
Nozawa, Kayo;O'Donoghue, Patrick;Gundllapalli, Sarath;Araiso, Yuhei;Ishitani, Ryuichiro;Umehara, Takuya;Soell, Dieter;Nureki, Osamu
吡咯赖氨酸（Pyl）是第22种天然氨基酸，由UAG基因编码，并通过独特的抑制基因tRNAPyl插入蛋白质中。甲烷菌科产生Pyl并表达允许在甲胺上生长的含Pyl的甲基转移酶。在两种细菌物种中也发现了同源甲基转移酶和Pyl生物合成和编码机制。Pyl编码由吡咯赖氨酰-tRNA合成酶（PylRS）维持，其催化Pyl-tRNAPyl的形成。Pyl不是最近才加入遗传密码的。PylRS已经存在于最后一个普遍的共同祖先中;然后它在利用甲胺作为能源的生物体中持续存在。最近的蛋白质工程努力将非规范氨基酸添加到遗传密码中。该技术依赖于“正交”tRNA合成酶：tRNA对的定向进化，其中工程化氨酰-tRNA合成酶（阿尔斯）特异性地且排他地用非规范氨基酸酰化正交tRNA。对于Pyl来说，自然进化过程在大约30亿年前发展了这样一个系统。当转化到大肠杆菌中时，巴氏甲烷八叠球菌PylRS和tRNAPyl在体内起正交对的作用。在这里，我们证明，Desulfitobacterium pylniense PylRS：tRNAPyl是一个正交对在体外和体内，并提出了这个正交对的晶体结构。PylRS：tRNAPyl的古老出现使得蛋白质和tRNA的独特结构特征得以进化。这些结构元素表现出一个复杂的，专门的阿尔斯：tRNA相互作用的表面高度不同于在任何其他已知的阿尔斯：tRNA复合物中观察到的那些;它是这种一般性质的基础正交性的分子基础。
Pyrrolysine (Pyl), the 22nd natural amino acid, is genetically encoded by UAG and inserted into proteins by the unique suppressor tRNAPyl. The Methanosarcinaceae produce Pyl and express Pyl-containing methyltransferases that allow growth on methylamines. Homologous methyltransferases and the Pyl biosynthetic and coding machinery are also found in two bacterial species. Pyl coding is maintained by pyrrolysyl-tRNA synthetase (PylRS), which catalyzes the formation of Pyl-tRNAPyl. Pyl is not a recent addition to the genetic code. PylRS was already present in the last universal common ancestor; it then persisted in organisms that utilize methylamines as energy sources. Recent protein engineering efforts added non-canonical amino acids to the genetic code. This technology relies on the directed evolution of an ‘orthogonal’ tRNA synthetase:tRNA pair in which an engineered aminoacyl-tRNA synthetase (aaRS) specifically and exclusively acylates the orthogonal tRNA with a non-canonical amino acid. For Pyl the natural evolutionary process developed such a system some 3 billion years ago. When transformed into Escherichia coli, Methanosarcina barkeri PylRS and tRNAPyl function as an orthogonal pair in vivo. Here we demonstrate that Desulfitobacterium hafniense PylRS:tRNAPyl is an orthogonal pair in vitro and in vivo, and present the crystal structure of this orthogonal pair. The ancient emergence of PylRS:tRNAPyl allowed for the evolution of unique structural features in both the protein and the tRNA. These structural elements manifest an intricate, specialized aaRS:tRNA interaction surface highly distinct from those observed in any other known aaRS:tRNA complex; it is this general property that underlies the molecular basis of orthogonality.
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DOI：
10.1016/j.bbrc.2008.07.074
发表时间：
2008-09-26
影响因子：
3.1
作者：
Lee, Marianne M.;Jiang, Ruisheng;Chan, Michael K.
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影响因子：
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作者：
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影响因子：
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作者：
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DOI：
10.1107/s0907444998003254
发表时间：
1998-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
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作者：
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