Ultrafast Protein Splicing is Common among Cyanobacterial Split Inteins: Implications for Protein Engineering
- 2 July 2012
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 134 (28) , 11338-11341
Abstract
We describe the first systematic study of a family of inteins, the split DnaE inteins from cyanobacteria. By measuring in vivo splicing efficiencies and in vitro kinetics, we demonstrate that several inteins can catalyze protein trans-splicing in tens of seconds rather than hours, as is commonly observed for this autoprocessing protein family. Furthermore, we show that when artificially fused, these inteins can be used for rapid generation of protein α-thioesters for expressed protein ligation. This comprehensive survey of split inteins provides indispensable information for the development and improvement of intein-based tools for chemical biology.Keywords
This publication has 18 references indexed in Scilit:
- Highly Efficient and More General cis- and trans-Splicing Inteins through Sequential Directed EvolutionJournal of Biological Chemistry, 2011
- Kinetic Control of One-Pot Trans-Splicing Reactions by Using a Wild-Type and Designed Split InteinAngewandte Chemie International Edition, 2011
- Modification of transmembrane and GPI-anchored proteins on living cells by efficient protein trans-splicing using the Npu DnaE inteinChemical Communications, 2011
- Biological Applications of Protein SplicingCell, 2010
- Backbone Dynamics and Global Effects of an Activating Mutation in Minimized Mtu RecA InteinsJournal of Molecular Biology, 2010
- Modulation of intein activity by its neighboring extein substratesProceedings of the National Academy of Sciences, 2009
- Traceless protein splicing utilizing evolved split inteinsProceedings of the National Academy of Sciences, 2009
- Solution structure of DnaE intein from Nostoc punctiforme: Structural basis for the design of a new split intein suitable for site‐specific chemical modificationFEBS Letters, 2009
- The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans‐splicing reactionFEBS Letters, 2009
- Highly efficient protein trans‐splicing by a naturally split DnaE intein from Nostoc punctiformeFEBS Letters, 2006