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Mutant pyrrolysyl-tRNA synthetase, and method for production of protein having non-natural amino acid integrated therein by using the same

机译:突变的吡咯基-tRNA合成酶,以及使用该酶合成整合有非天然氨基酸的蛋白质的方法

摘要

Method for incorporating a lysine derivative (particularly an Nε-benzyloxycarbonyl-lysine (Z-Lys) derivative) having useful functional group such as heavy atom, selenium, reactive functional group, fluorescent group or crosslinker, which is suitable as a non-natural amino acid, into a desired protein in a site-specific manner. A mutant pyrrolysyl-tRNA synthetase has substitution of at least one amino acid residue selected from tyrosine residue at position 306, leucine residue at position 309 and cysteine residue at position 348 each constituting a pyrrolysine-binding site in the amino acid sequence for pyrrolysyl-tRNA synthetase of SEQ ID NO:2. The substitution of the amino acid residue is: of tyrosine residue at position 306 by glycine or alanine residue, of leucine residue at position 309 by glycine or alanine residue, and/or of a cysteine residue at position 348 by valine, serine or alanine residue.
机译:掺入具有有用的官能团如重原子,硒,反应性官能团,荧光基团或交联剂的赖氨酸衍生物(特别是N ε-苄氧基羰基赖氨酸(Z-Lys)衍生物)的方法,以位点特异性的方式适合作为所需蛋白的非天然氨基酸。突变的吡咯赖氨酸-tRNA合成酶具有至少一个氨基酸残基的取代,所述氨基酸残基选自306位的酪氨酸残基,309位的亮氨酸残基和348位的半胱氨酸残基,它们各自构成吡咯赖氨酸-tRNA的氨基酸序列中的吡咯赖氨酸结合位点。 SEQ ID NO:2的合成酶。氨基酸残基的取代是:306位的酪氨酸残基被甘氨酸或丙氨酸残基取代,309位的亮氨酸残基被甘氨酸或丙氨酸残基取代,和/或缬氨酸,丝氨酸或丙氨酸残基在348位的半胱氨酸残基取代。 。

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