Three-Segment Protein Labeling Using a Highly Efficient and Cysteine-Less Split Intein Identified with Computational Prediction of Aggregation Properties.

ABSTRACT: Split inteins are versatile tools in protein engineering. We envisaged a new tandem protein trans-splicing (PTS) scheme to assemble a protein from three segments, of which each can be individually treated with regard to its cysteine oxidation or chemical labeling status. However, only a single highly efficient cysteine-less split intein has been reported so far. Split intein activities are currently not predictable and require time-consuming biochemical characterizations. We aimed to accelerate the discovery of novel split inteins with high splicing efficiency by computational sequence analysis. Inspired by our previous finding that linked reduced splicing efficiency of characterized split intein fragments to soluble, β-sheet-rich amyloid-like aggregates, we confirmed the inverse correlati

SUBMITTER: Humberg C 

PROVIDER: S-EPMC12707363 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature