METHODS FOR ENZYMATIC PEPTIDE LIGATION
    1.
    发明公开

    公开(公告)号:US20240263210A1

    公开(公告)日:2024-08-08

    申请号:US18465824

    申请日:2023-09-12

    CPC classification number: C12P21/02 C07K14/415 C12N9/93

    Abstract: The present invention relates to identification and utilization of a novel structural feature governing efficient peptide ligation of OaAEP1 and a method of forming a peptide of Formula (I) by ligating a first peptide of Formula (II) to a second peptide of Formula (III) by enzymatically cleaving the bond between “Asx” and “Xaa3” in the first peptide of Formula (II) and ligating the fragment P1-Asx of the first peptide via its C-terminus to the N-terminus of the second peptide of Formula (III) to form a ligated peptide of Formula (I), wherein the enzymatic cleavage and ligation reaction is catalyzed by a peptide ligase having the activity of OaAEP1 Cys247Ala (SEQ ID NO:1) under conditions suitable for said cleavage and ligation reaction. Further encompassed are a method of preparing a dimer, oligomer, or multimer of one or more peptides of interest and a method of modifying or tagging the surface of a target cell by one or more peptides of interest catalyzed by the peptide ligase having the activity of OaAEP1 Cys247Ala (SEQ ID NO:1). Also encompassed in the invention are the ligated peptides and/or tagged target cells obtainable according to any of the methods of the invention, the peptide ligase OaAEP1 Cys247Ala having the amino acid sequence of SEQ ID NO:1 and other peptide ligases having the activity thereof, as well as kits comprising any of said peptide ligases, in particular OaAEP1 Cys247Ala (SEQ ID NO:1).

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