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Generation of engineered recombinant hepatocyte growth factor cleaved and activated by Genenase I
- Source :
-
Journal of Biotechnology . Feb2008, Vol. 133 Issue 4, p478-485. 8p. - Publication Year :
- 2008
-
Abstract
- Abstract: Hepatocyte growth factor (HGF) is biosynthesized as a biologically inactive, single-chain form (pro-HGF). Its activation is associated with cleavage at Arg494-Val495 into a two-chain mature form composed of disulfide-linked α- and β-chains. Because serum is a major source of HGF activator (the predominant serine protease responsible for the processing of pro-HGF), serum-free production of recombinant, two-chain HGF had not been established. In this study, to enable serum-free production of two-chain HGF, we generated engineered human pro-HGFs that can be specifically cleaved and activated by Genenase I. Since Genenase I specifically cleaves the C-terminus of the His-Tyr sequence, which does not exist in human HGF, Arg494 (the C-terminus of the HGF α-chain) was replaced by His-Tyr, Ala-Ala-His-Tyr, Pro-Gly-His-Tyr, or Pro-Gly-Ala-Ala-His-Tyr. Genenase I cleaved engineered pro-HGFs specifically at the replaced amino acid sequences, forming a disulfide-linked two-chain form. The cleavage was most efficient in the case of the Pro-Gly-Ala-Ala-His-Tyr sequence, and cleaved HGFs displayed biological activities identical to those of wild-type HGF. Considering a potential medical application of HGF, the present technique is valuable because it enables the production of recombinant, two-chain HGF entirely without serum and extends the choice of host cells and organisms for recombinant production. [Copyright &y& Elsevier]
- Subjects :
- *LIVER cells
*GROWTH factors
*PROTEOLYTIC enzymes
*HYDROLASES
Subjects
Details
- Language :
- English
- ISSN :
- 01681656
- Volume :
- 133
- Issue :
- 4
- Database :
- Academic Search Index
- Journal :
- Journal of Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 29375576
- Full Text :
- https://doi.org/10.1016/j.jbiotec.2007.11.006