Biological Activity of Brain-derived Neurotrophic Factor with Mismatched Disulfide Linkages Produced by Escherichia coli

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 1996-06, Vol.60 (6), p.971-974
• Main Authors: Shimizu, Norio, Fukuzono, Shinichi, Fujimori, Kiyoshi, Park, Kyungmoon, Takeshita, Tomoko, Ishibashi, Tadashi
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; BDNF produced by Escherichia coli; BDNF with mismatched disulfide linkages; biological activity of BDNF; Biological and medical sciences; Biotechnology; Brain-Derived Neurotrophic Factor; brain-derived neurotrophic factor (BDNF); Chick Embryo; Cloning, Molecular; Disulfides - metabolism; Escherichia coli; Fundamental and applied biological sciences. Psychology; Nerve Tissue Proteins - chemistry; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neurons - drug effects; neurotrophic factor; Protein hormones. Growth factors. Cytokines; Proteins; Rats; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1271/bbb.60.971

E. coli cells harboring an expression plasmid with a brain-derived neurotrophic factor (BDNF) gene obtained from rat BDNF cDNA were sonicated and centrifuged to obtain a precipitate containing BDNF. Ten different proteins of BDNF were purified from the precipitate and the three disulfide linkages of six proteins were identified. Those disulfide structures were different from that of authentic BDNF and the biological activities of BDNF with mismatched disulfide linkages (EC 50 of 2 to 15ng/ml) were much lower than that of authentic BDNF (EC 50 of 30pg/ml). The BDNF with mismatched disulfide linkages inhibited the biological activity of authentic BDNF.