Process for the specific cleavage of protein sequences from prot

Details Process for the specific cleavage of protein sequences from prot Process for the specific cleavage of protein sequences from prot

1982-07-06

1985-09-24

Kepplinger, Esther M.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

435 70, 4351723, 935 47, 935 51, C12P 2106, C12P 2102, C12N 1500

Patent

active

045433291

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-Zxy-bond by treating the protein with postproline dipeptidylaminopeptidase (PPDA), thereby producing protein having the sequence Zxy-Uvw-Pro at its N-terminal end, followed by cleaving the Zxy acid from the resultant protein by treating the latter with leucine aminopeptidase.

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