Please use this identifier to cite or link to this item:
Scopus Web of Science® Altmetric
Type: Journal article
Title: Introduction of spacer peptides N-terminal to a cleavage recognition motif in recombinant fusion proteins can improve site-specific cleavage
Author: Polyak, S.
Forsberg, G.
Forbes, B.
McNeil, K.
Aplin, S.
Wallace, J.
Citation: Protein Engineering Design and Selection, 1997; 10(6):615-619
Publisher: Oxford University Press
Issue Date: 1997
ISSN: 0269-2139
Statement of
Steve W Polyak, Göran Forsberg, Briony E Forbes, Kerrie A McNeil, Sally E Aplin, and John C Wallace
Abstract: To improve site-specific cleavage of a methionyl porcine growth hormone [[Met1]-pGH(1-46)-IGF-II] fusion protein by the enzyme H64A subtilisin, a series of flexible, unstructured spacer peptides were introduced N-terminal to the cleavage site. When enzymatic digestion preceded refolding of the fusion proteins, IGF-II could only be liberated from substrates which contained spacer peptides. Compared with the parent construct, the yield of IGF-II from refolded fusion proteins containing spacers was improved up to two-fold. Furthermore, this cleavage rate was improved by removing a competing protease recognition motif from the fusion partner. These data show that fusion partners can influence site-specific proteolysis of fusion proteins. Introduction of flexible spacers between the moieties can alleviate these interactions.
Keywords: Animals
Growth Hormone
Insulin-Like Growth Factor II
Protein Sorting Signals
Recombinant Proteins
Recombinant Fusion Proteins
Mutagenesis, Site-Directed
Amino Acid Sequence
Protein Folding
Molecular Sequence Data
DOI: 10.1093/protein/10.6.615
Appears in Collections:Aurora harvest 7
Biochemistry publications

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.