{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"omics_type":["Unknown"],"volume":["6(2)"],"submitter":["Cui T"],"pubmed_abstract":["Site-specific protein modification enables precise functionalization for applications ranging from live-cell imaging to therapeutic development. Although N-terminal cysteine (N-Cys)-based modification exhibits excellent specificity, naturally occurring N-Cys is rare, and existing methods are usually limited by factors such as narrow substrate scopes, incompatibility with inclusion bodies, or residual exogenous sequences. Here, we report a highly efficient and versatile method for generating free N-Cys proteins through \"Lys-Leu\" (K-L) motif-assisted <i>Oa</i>AEP1-catalyzed cleavage. This approach enhances cleavage efficiency across all 20 natural amino acids at the P2' position, accommodates diverse substrates (including challenging inclusion body proteins), and enables streamlined one-pot "],"journal":["JACS Au"],"pagination":["943-954"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC12933375"],"repository":["biostudies-literature"],"pubmed_title":["\"Lys-Leu\" Motif Empowers Expedient &lt;i&gt;Oa&lt;/i&gt;AEP1-Catalyzed Generation of N‑Terminal Cysteine Recombinant Proteins for Bioconjugation and Semisynthesis."],"pmcid":["PMC12933375"],"pubmed_authors":["Li J","Liu N","Li YM","Cui T","Sun J","Chu GC"],"additional_accession":[]},"is_claimable":false,"name":"\"Lys-Leu\" Motif Empowers Expedient &lt;i&gt;Oa&lt;/i&gt;AEP1-Catalyzed Generation of N‑Terminal Cysteine Recombinant Proteins for Bioconjugation and Semisynthesis.","description":"Site-specific protein modification enables precise functionalization for applications ranging from live-cell imaging to therapeutic development. Although N-terminal cysteine (N-Cys)-based modification exhibits excellent specificity, naturally occurring N-Cys is rare, and existing methods are usually limited by factors such as narrow substrate scopes, incompatibility with inclusion bodies, or residual exogenous sequences. Here, we report a highly efficient and versatile method for generating free N-Cys proteins through \"Lys-Leu\" (K-L) motif-assisted <i>Oa</i>AEP1-catalyzed cleavage. This approach enhances cleavage efficiency across all 20 natural amino acids at the P2' position, accommodates diverse substrates (including challenging inclusion body proteins), and enables streamlined one-pot ","dates":{"release":"2026-01-01T00:00:00Z","publication":"2026 Feb","modification":"2026-07-16T22:57:44.428Z","creation":"2026-07-12T03:09:54.396Z"},"accession":"S-EPMC12933375","cross_references":{"pubmed":["41755832"],"doi":["10.1021/jacsau.5c01395"]}}