An Atypical Mechanism of Split Intein Molecular Recognition and Folding

Journal of the American Chemical Society
Adam J StevensTom W Muir

Abstract

Split inteins associate to trigger protein splicing in trans, a post-translational modification in which protein sequences fused to the intein pair are ligated together in a traceless manner. Recently, a family of naturally split inteins has been identified that is split at a noncanonical location in the primary sequence. These atypically split inteins show considerable promise in protein engineering applications; however, the mechanism by which they associate is unclear and must be different from that of previously characterized canonically split inteins due to unique topological restrictions. Here, we use a consensus design strategy to generate an atypical split intein pair (Cat) that has greatly improved activity and is amenable to detailed biochemical and biophysical analysis. Guided by the solution structure of Cat, we show that the association of the fragments involves a disorder-to-order structural transition driven by hydrophobic interactions. This molecular recognition mechanism satisfies the topological constraints of the intein fold and, importantly, ensures that premature chemistry does not occur prior to fragment complementation. Our data lead a common blueprint for split intein complementation in which localized s...Continue Reading

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Citations

Oct 16, 2019·Proceedings of the National Academy of Sciences of the United States of America·Maniraj BhagawatiHenning D Mootz
May 31, 2020·Nature Chemistry·Antony J BurtonTom W Muir
May 20, 2020·Proceedings of the National Academy of Sciences of the United States of America·Antony J BurtonTom W Muir
Nov 28, 2019·Chemical Reviews·Robert E Thompson, Tom W Muir
Aug 17, 2019·Journal of the American Chemical Society·Josef A GramespacherTom W Muir
Nov 24, 2021·Journal of Peptide Science : an Official Publication of the European Peptide Society·Huasong AiJia-Bin Li

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