title: Proteinase-catalyzed conversion of a substance P-precursor peptide.
abstract: The protease-catalyzed conversion of peptides and proteins produced by recombinant DNA technology is a promising method for large-scale production of peptides including those with non-proteinogenic structural elements. As a model system we have investigated the proteinase-catalyzed modification of a chemically synthesized substance P-precursor. In the precursor peptide the residues of substance P(1-8) were flanked by tripeptide linkers on both sides. In the first step the C-terminal tripeptide amide was replaced by the authentic C-terminal tripeptide amide of substance P via alpha-chymotrypsin-catalyzed transpeptidation. The enzyme simultaneously attacks two peptide bonds in the precursor molecule leading to the formation of several side-products. The desired peptide was obtained with 25% yield. In the second step the other tripeptide linker was selectively and almost quantitatively removed from the N-terminus of the precursor via trypsin-catalyzed hydrolysis. This study demonstrates that substance P can be obtained from an engineered protein by proteinase-catalyzed processing.
authors.affiliation: GBF (Gesellschaft für Biotechnologische Forschung mbH), Braunschweig, Germany.
authors.forename: V
authors.initials: V
authors.lastname: Schellenberger
authors.forename: W
authors.initials: W
authors.lastname: Tegge
authors.forename: K D
authors.initials: KD
authors.lastname: Klöppel
authors.forename: R
authors.initials: R
authors.lastname: Frank
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002918:Chymotrypsin / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism*; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D013373:Substance P / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*
chemical_list: D011498:Protein Precursors; D013373:Substance P; D010450:Endopeptidases; D002918:Chymotrypsin