Generation of engineered recombinant hepatocyte growth factor cleaved and activated by Genenase I

Journal of Biotechnology
Daichika HayataToshikazu Nakamura

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 alpha- and beta-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 alpha-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 v...Continue Reading

References

Dec 30, 1992·Biochemical and Biophysical Research Communications·K MizunoT Nakamura
Jan 1, 1989·Proteins·P CarterJ A Wells
Sep 15, 1989·Biochemical and Biophysical Research Communications·K MiyazawaK Takahashi
Aug 16, 1984·Biochemical and Biophysical Research Communications·T NakamuraA Ichihara
Nov 21, 1997·Biochemical and Biophysical Research Communications·K Matsumoto, T Nakamura
Oct 28, 1998·Trends in Cell Biology·C Birchmeier, E Gherardi
May 31, 2001·Kidney International·K Matsumoto, T Nakamura
Dec 17, 2002·Clinica Chimica Acta; International Journal of Clinical Chemistry·Hiroshi Funakoshi, Toshikazu Nakamura
Apr 9, 2004·Proceedings of the National Academy of Sciences of the United States of America·Chang-Goo HuhSnorri S Thorgeirsson
Jul 14, 2004·Proceedings of the National Academy of Sciences of the United States of America·Malgorzata BorowiakCarmen Birchmeier
Feb 8, 2005·The Biochemical Journal·Kazuhiro FukutaToshikazu Nakamura
Mar 21, 2007·Proceedings of the National Academy of Sciences of the United States of America·Daniel KirchhoferRobert A Lazarus
May 1, 2007·Regenerative Medicine·Shinya Mizuno, Toshikazu Nakamura

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.