Examining the structural and chemical flexibility of the active site base, Lys-258, of Escherichia coli aspartate aminotransferase by replacement with unnatural amino acids.
Examining the structural and chemical flexibility of the active site base, Lys-258, of Escherichia coli aspartate aminotransferase by replacement with unnatural amino acids.
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通过用非天然氨基酸替换来检查大肠杆菌天冬氨酸转氨酶活性位点碱基 Lys-258 的结构和化学灵活性。
DOI：
10.1021/bi00038a028
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发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Kirsch,JF
中科院分区：
文献类型：
--
作者：
Gloss,LM;Kirsch,JF
方法K258 Cq的定点突变已在别处描述[参见Gloss和Kirsch（1995 a）及其引用的参考文献]，K258 Eq通过相同的方法制备。根据Gloss et al.（1992），进行了以下简化：将硫酸铵沉淀在G-100柱上的色谱法替换为对20 mM磷酸钾缓冲液（pH 7.5）、5 mM磷酸钾缓冲液（pH 7.5）进行透析。
MethodsSite-directed mutagenesis of K258Cq has been described elsewhere [see Gloss and Kirsch (1995a) and references cited therein], K258Eq was prepared by the same method. Purification and quantification of WT and mutant AATases were performed according to Gloss et al.(1992), with the following simplification: chromatography of the ammonium sulfate pellet over a G-100 column was replaced by dialysis against 20 mM potassium phosphate buffer, pH 7.5, 5 mM