An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein.
An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein.
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DOI：
10.1038/s41598-021-91090-w
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发表时间：
2021-06-03
影响因子：
4.6
通讯作者：
Mills KV
中科院分区：
文献类型：
--
作者：
Williams JE;Jaramillo MV;Li Z;Zhao J;Wang C;Li H;Mills KV
蛋白质剪接是一种翻译后过程，内含肽通过该过程催化其自身从侧翼多肽或外显肽切除，伴随外显肽连接。许多内含肽具有巢式归巢核酸内切酶结构域，其促进它们传播到无内含肽的等位基因中，而其他内含肽缺乏归巢核酸内切酶（HEN）并且被称为迷你内含肽。中断Pyrococcus horikoshii的DNA PolII的迷你内含肽具有代替HEN结构域的接头区域，其比来自深海Pyrococcus abyssi的密切相关内含肽中的接头短。堀越毕赤酵母PolII内含肽需要更高的温度以获得催化活性，并且对热稳定蛋白酶嗜热菌蛋白酶的消化更稳定，表明它比深海毕赤酵母内含肽更刚性。我们解决了内含肽前体的晶体结构，揭示了结构域交换的二聚体。作为结构域交换二聚体发现的内含肽已显示促进内含肽介导的蛋白质选择性剪接，但解析的P. horikoshii PolII内含肽结构具有不太可能具有催化活性的活性位点。
Protein splicing is a post-translational process by which an intein catalyzes its own excision from flanking polypeptides, or exteins, concomitant with extein ligation. Many inteins have nested homing endonuclease domains that facilitate their propagation into intein-less alleles, whereas other inteins lack the homing endonuclease (HEN) and are called mini-inteins. The mini-intein that interrupts the DNA PolII of Pyrococcus horikoshii has a linker region in place of the HEN domain that is shorter than the linker in a closely related intein from Pyrococcus abyssi. The P. horikoshii PolII intein requires a higher temperature for catalytic activity and is more stable to digestion by the thermostable protease thermolysin, suggesting that it is more rigid than the P. abyssi intein. We solved a crystal structure of the intein precursor that revealed a domain-swapped dimer. Inteins found as domain swapped dimers have been shown to promote intein-mediated protein alternative splicing, but the solved P. horikoshii PolII intein structure has an active site unlikely to be catalytically competent.
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