6DSL

Consensus engineered intein (Cat) with atypical split site

6DSL の概要

• エントリーDOI: 10.2210/pdb6dsl/pdb
• NMR情報: BMRB: 30480
• 分子名称: Consensus engineered intein CatN, Consensus engineered intein CatC (2 entities in total)
• 機能のキーワード: intein protein, trans splicing, consensus engineering, protein engineering, splicing
• 由来する生物種: Enterobacteria phage T7; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 17295.47

構造登録者

Sekar, G.,Stevens, A.J.,Muir, T.W.,Cowburn, D. (登録日: 2018-06-14, 公開日: 2018-09-19, 最終更新日: 2024-05-01)

主引用文献

Stevens, A.J.,Sekar, G.,Gramespacher, J.A.,Cowburn, D.,Muir, T.W.
An Atypical Mechanism of Split Intein Molecular Recognition and Folding.
J. Am. Chem. Soc., 140:11791-11799, 2018

PubMed 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 structural rearrangements are used to drive folding and regulate protein-splicing activity.

PubMed: 30156841
DOI: 10.1021/jacs.8b07334

実験手法

SOLUTION NMR