Research Program in Structural Biology and Biophysics, Institute of Biotechnology, University of Helsinki. P.O. Box 65, Helsinki, FIN-00014, Finland.
FEBS Lett. 2011 Nov 4;585(21):3471-7. doi: 10.1016/j.febslet.2011.10.005. Epub 2011 Oct 10.
Protein splicing catalyzed by inteins has enabled various biotechnological applications such as protein ligation. Successful applications of inteins are often limited by splicing efficiency. Here, we report the comparison of protein splicing between 20 different inteins from various organisms in identical contexts to identify robust inteins with foreign exteins. We found that RadA intein from Pyrococcus horikoshii and an engineered DnaB intein from Nostoc punctiforme demonstrated an equally efficient splicing activity to the previously reported highly efficient DnaE intein from Nostoc punctiforme. The newly identified inteins with efficient cis-splicing activity can be good starting points for the further development of new protein engineering tools.
蛋白质剪接由内肽酶催化，已实现了各种生物技术应用，如蛋白质连接。内肽酶的成功应用通常受到剪接效率的限制。在这里，我们在相同的环境下比较了来自不同生物体的 20 种不同内肽酶的蛋白质剪接，以鉴定具有外源外肽酶的稳健内肽酶。我们发现来自 Pyrococcus horikoshii 的 RadA 内肽酶和来自 Nostoc punctiforme 的工程化 DnaB 内肽酶与之前报道的来自 Nostoc punctiforme 的高效 DnaE 内肽酶具有同样高效的剪接活性。具有高效顺式剪接活性的新鉴定内肽酶可以成为进一步开发新蛋白质工程工具的良好起点。
