Process for the specific cleavage of protein sequences from proteins

US4543329A · kind A · utility

Assignee

• SCHERING CORPORATION · US

Inventors

• Joachim Daum · Berlin, DE
• Gerhard Siewert · Berlin, DE
• Michael Topert · Berlin, DE
• Hartmut Seliger · Neu-Ulm, DE

Key dates

• Filing date: Jul 6, 1982
• Grant date: Sep 24, 1985
• Expiry date: Jul 6, 2002

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10S930/31
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A process for cleavage of a protein (a) sequence from a protein containing the tetrapeptide sequence Pro-Xyz-Gly-Pro, wherein Xyz can be any desired amino acid, and reacting the protein with a collagenase whereby the Xyz-Gly bond is selectively cleaved, thereby producing a protein with the sequence Gly-Pro at its N-terminal end; or (b) containing the sequence Met-Pro; and subsequently selectively cleaving the glycine or methionine residue from the N-terminal end of the product protein with an aminoacylproline aminopeptidase and selectively cleaving the proline residue therefrom with a proline aminopeptidase; or subsequently selectively cleaving the Gly-Pro or Met-Pro residue from the N-terminal end of the produced protein using a postproline dipeptidylaminopeptidase. In a special embodiment, the protein which is split contains the sequence -Zxy-Uvw-Pro- on the carboxy end of the sequence Pro-Xyz-Gly-Pro, wherein Zxy and Uvw are independently any natural amino acid in the genetic code except Pro, whereby after the collagenase reaction, there is produced a protein with the sequence Gly-Pro-Zxy-Uvw-Pro at the N-terminal end. This is split by subsequently selectively cleaving the Pro-Z…