The crystal structure of the naturally split gp41-1 intein guides the engineering of orthogonal split inteins from cis-splicing inteins
The crystal structure of the naturally split gp41-1 intein guides the engineering of orthogonal split inteins from cis-splicing inteins
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DOI：
10.1111/febs.15113
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发表时间：
2019-11-19
期刊：
影响因子：
5.4
通讯作者：
Iwai, Hideo
中科院分区：
文献类型：
--
作者：
Beyer, Hannes M.;Mikula, Kornelia M.;Iwai, Hideo
由分裂内含子催化的蛋白质反式剪接作为一种蛋白质工程工具越来越有用。我们解决了来自自然分裂的gp41-1内含子的融合变体的1.0埃分辨率的晶体结构，这是先前从环境元基因组序列数据中确定的。125个残基的gp41-1内含子的结构显示出在Hedgehog/Intein超家族中常见的紧凑的伪C2对称性，在分裂的N-末端和C-末端的内含子片段之间存在广泛的电荷-电荷相互作用，这在自然产生的分裂内含子中是常见的。我们通过将类似的电荷网络工程到顺式剪接内含子的相同区域，成功地创建了正交分裂内含子。这一策略可以应用于从其他更普遍的顺式剪接内含子中创建新的自然类分裂内含子。数据库结构数据可在RCSB蛋白质数据库中的登录号下获得。
Protein trans-splicing catalyzed by split inteins has increasingly become useful as a protein engineering tool. We solved the 1.0 angstrom-resolution crystal structure of a fused variant from the naturally split gp41-1 intein, previously identified from environmental metagenomic sequence data. The structure of the 125-residue gp41-1 intein revealed a compact pseudo-C2-symmetry commonly found in the Hedgehog/Intein superfamily with extensive charge-charge interactions between the split N- and C-terminal intein fragments that are common among naturally occurring split inteins. We successfully created orthogonal split inteins by engineering a similar charge network into the same region of a cis-splicing intein. This strategy could be applicable for creating novel natural-like split inteins from other, more prevalent cis-splicing inteins. Database Structural data are available in the RCSB Protein Data Bank under the accession number .