Facile expression of proteins with desired N-terminal amino acid via an engineered cysteine protease domain.

ABSTRACT: The removal of N-terminal methionine (Met) or fused N-terminal purification tags to expose the first amino acid of the target protein is critically important for studying the potential regulatory role of the N-terminal residue. However, current tag-removal approaches typically rely on enzymatic cleavage or require harsh reaction conditions. Here, we report a strategy for expressing proteins of interest (POI) with custom N-terminal amino acids by introducing an engineered cysteine protease domain (CPD) tag at the N-terminus. The cleavable tag is chemically triggered by inositol hexakisphosphate (InsP₆), enabling precise generation of proteins with a user-defined N-terminus. Through systemic design and engineering of the N-terminal CPD tag, we successfully achieved POI variants wi

SUBMITTER: Zou X 

PROVIDER: S-EPMC12329031 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature