A new strategy for the synthesis of glycoproteins.
A new strategy for the synthesis of glycoproteins.
复制标题
糖蛋白合成的新策略。
DOI：
10.1126/science.1089509
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Schultz,PeterG
中科院分区：
文献类型：
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作者：
Zhang,Zhiwen;Gildersleeve,Jeff;Yang,Yu-Ying;Xu,Ran;Loo,JosephA;Uryu,Sean;Wong,Chi-Huey;Schultz,PeterG
蛋白质的翻译后修饰调控许多生物过程，包括新陈代谢、信号转导和基因表达。然而，与产生选择性修饰蛋白质的同质群体相关的合成挑战阻碍了对这些修饰对蛋白质结构和功能影响的详细研究。在这里，我们报告了一种选择性糖基化蛋白质的共翻译合成方法，其中修饰的氨基酸是基因编码的。我们发现，在特定位置含有β-N-乙酰氨基葡萄糖(GlcNAc)-丝氨酸的肌红蛋白可以在大肠杆菌中表达，产量高，保真度高。β-GlcNAc部分可被糖结合蛋白识别，或随后被半乳糖基转移酶修饰以建立更复杂的碳水化合物。这种方法应该普遍适用于其他翻译后修饰，如蛋白质磷酸化、乙酰化和甲基化。
Posttranslational modifications of proteins regulate many biological processes, including metabolism, signal transduction, and gene expression. The synthetic challenges associated with generating homogeneous populations of selectively modified proteins, however, have hindered detailed studies of the effects of these modifications on protein structure and function. Here, we report an approach to the cotranslational synthesis of selectively glycosylated proteins in which the modified amino acid is genetically encoded. We show that myoglobin containing β–N-acetylglucosamine (GlcNAc)–serine at a defined position can be expressed inEscherichia coliin good yield and with high fidelity. The β-GlcNAc moiety can be recognized by a saccharide-binding protein, or subsequently modified with a galactosyltransferase to build more complex carbohydrates. This approach should be generally applicable to other posttranslational modifications such as protein phosphorylation, acetylation, and methylation.