@article{oai:kanazawa-u.repo.nii.ac.jp:00027388,
 author = {松本, 邦夫 and マツモト, クニオ and Hayata, Daichika and Fukuta, Kazuhiro and 松本, 邦夫 and マツモト, クニオ and Adachi, Eri and Hanada, Keigo and Adachi, Kiichi and Nakamura, Toshikazu},
 issue = {4},
 journal = {Journal of Biotechnology},
 month = {Feb},
 note = {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. © 2007 Elsevier B.V. All rights reserved., 金沢大学がん研究所附属分子標的がん医療研究開発センター},
 pages = {478--485},
 title = {Generation of engineered recombinant hepatocyte growth factor cleaved and activated by Genenase I},
 volume = {133},
 year = {2008}
}