Structure-based engineering and comparison of novel split inteins for protein ligation.
Structure-based engineering and comparison of novel split inteins for protein ligation.
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基于结构的工程和用于蛋白质连接的新型分裂蛋白的比较。
DOI：
10.1039/c4mb00021h
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发表时间：
2014-05
影响因子：
--
通讯作者：
Iwaï H
中科院分区：
文献类型：
--
作者：
Aranko AS;Oeemig JS;Zhou D;Kajander T;Wlodawer A;Iwaï H
蛋白质剪接是一种自催化过程，涉及蛋白质内部结构域（内含肽）的自我切除，以及伴随的两个侧翼序列（外蛋白）与肽键的连接。蛋白质剪接也可以通过天然分裂的内含肽或人工分裂的内含肽反式发生，将两条不同多肽链上的内含肽连接成一条多肽链。蛋白质反式剪接可以通过用其他蛋白质序列替换天然外显肽序列而在外源环境中起作用。使用蛋白质反式剪接的蛋白质连接日益成为生物技术应用如蛋白质半合成、片段同位素标记和体内蛋白质工程的有用工具。然而，只有少数分裂内含肽已成功地应用于蛋白质连接。天然分裂内含肽已被广泛使用，但它们彼此交叉反应，限制了它们的应用多片段连接。基于包含两个新确定的内含肽结构的三维结构，我们从四个高效顺式剪接的内含肽中衍生出21个新的分裂内含肽，以开发适合于蛋白质连接的新型分裂内含肽。我们系统地比较了24个分裂内含肽的反式剪接，并测试了它们之间的交叉活性，以确定正交分裂内含肽片段，可用于化学生物学和生物技术应用。
Protein splicing is an autocatalytic process involving self-excision of an internal protein domain, the intein, and concomitant ligation of the two flanking sequences, the exteins, with a peptide bond. Protein splicing can also take place in trans by naturally split inteins or artificially split inteins, ligating the exteins on two different polypeptide chains into one polypeptide chain. Protein trans-splicing could work in foreign contexts by replacing the native extein sequences with other protein sequences. Protein ligation using protein trans-splicing increasingly becomes a useful tool for biotechnological applications such as semi-synthesis of proteins, segmental isotopic labeling, and in vivo protein engineering. However, only a few split inteins have been successfully applied for protein ligation. Naturally split inteins have been widely used, but they are cross-reactive to each other, limiting their applications to multiple-fragment ligation. Based on the three-dimensional structures including two newly determined intein structures, we derived 21 new split inteins from four highly efficient cis-splicing inteins, in order to develop novel split inteins suitable for protein ligation. We systematically compared trans-splicing of 24 split inteins and tested the cross-activities among them to identify orthogonal split intein fragments that could be used in chemical biology and biotechnological applications.
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影响因子：
14.8
作者：
Aranko, A. Sesilja;Oeemig, Jesper S.;Iwai, Hideo
通讯作者：
Iwai, Hideo
影响因子：
2.9
作者：
Brenzel, S;Kurpiers, T;Mootz, HD
通讯作者：
Mootz, HD
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
64.5
作者：
Hall, TMT;Porter, JA;Leahy, DJ
通讯作者：
Leahy, DJ
影响因子：
3.5
作者：
Ellila, Simo;Jurvansuu, Jaana Maria;Iwai, Hideo
通讯作者：
Iwai, Hideo