The aim of this work was the development of a method for N-terminal, regiospecific peptide and protein modification using the reverse proteolysis reaction. The conditions for the segment coupling were optimized on a model peptide (Carbohydrate Structure Mimicking Peptide) with IgA protease and than applied to proteins. On the N-terminus of two recombinant proteins (with site directed mutagenesis modified anticalin FluA: IgAP1 and IgAP2) a tetra peptide (PRPP) with an artificial protecting group (Boc) was successfully connected. The structures of the modified proteins were determined with MALDI-TOF-MS and Edman Degradation. This new method for protein modification enables selective (regio-, chemo- and stereo selective), mild and very varied in vitro derivatization of native proteins and opens new application fields, e.g. analysis, separation, cleaning, immobilization, quantification of proteins, protein engineering, clinical studies, production of protein fusions, protein semi synthesis and peptide segment coupling.
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The aim of this work was the development of a method for N-terminal, regiospecific peptide and protein modification using the reverse proteolysis reaction. The conditions for the segment coupling were optimized on a model peptide (Carbohydrate Structure Mimicking Peptide) with IgA protease and than applied to proteins. On the N-terminus of two recombinant proteins (with site directed mutagenesis modified anticalin FluA: IgAP1 and IgAP2) a tetra peptide (PRPP) with an artificial protecting group...
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