title: Efficient cleavage by alpha-thrombin of a recombinant fused protein which contains insulin-like growth factor I.
abstract: The gene for insulin-like growth factor I (IGF-I) was constructed from chemically synthesized deoxyoligonucleotides and expressed in Escherichia coli, under the control of a trp promoter, as a set of fusion proteins which were connected with a portion of human growth hormone through the recognition sequence for a sequence-specific protease, either blood coagulation factor Xa or alpha-thrombin. Upon induction with 3-indoleacrylic acid, fusion proteins accumulated with a yield of 10-30% of the total protein. A fusion protein connected through a tetradecapeptide (Asp-Asp-Pro-Pro-Thr-Val-Glu-Leu-Gln-Gly-Leu-Val-Pro-Arg) was efficiently and correctly cleaved by alpha-thrombin, and the purified IGF-I possessed somatomedin-like activity, as determined by the enhancement of sulfation of glycosaminoglycans in cultured costal chondrocytes from rabbits.
authors.affiliation: Faculty of Pharmaceutical Sciences, Osaka University, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Nishikawa
authors.forename: K
authors.initials: K
authors.lastname: Yanase
authors.forename: T
authors.initials: T
authors.lastname: Tokunaga-Doi
authors.forename: K
authors.initials: K
authors.lastname: Kodama
authors.forename: H
authors.initials: H
authors.lastname: Gomi
authors.forename: S
authors.initials: S
authors.lastname: Uesugi
authors.forename: E
authors.initials: E
authors.lastname: Ohtsuka
authors.forename: Y
authors.initials: Y
authors.lastname: Kato
authors.forename: F
authors.initials: F
authors.lastname: Suzuki
authors.forename: M
authors.initials: M
authors.lastname: Ikehara
mesh_terms: D001681:Biological Assay; D011088:DNA Ligases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015951:Factor Xa; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D010957:Plasmids; D011863:Radioimmunoassay; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D012697:Serine Endopeptidases / Q000378:metabolism; D013002:Somatomedins / Q000378:metabolism*; D013917:Thrombin / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D013917:Thrombin; D015951:Factor Xa; D011088:DNA Ligases