Synthesis of new peptides with prolactin-releasing activity by a combination of recombinant DNA technology and a cysteine-specific cyanylation reaction

Chemical and Pharmaceutical Bulletin

Volumn 46, Issue 9, 1998, Pages 1490-1492

  Nishimura, Osamu ^a   Moriya, Takeo ^a   Suenaga, Masato ^a   Tanaka, Yoko ^a   Itoh, Takashi ^a   Koyama, Nobuyuki ^a   Fujii, Ryo ^a   Hinuma, Shuji ^a   Kitada, Chieko ^a   Fujino, Masahiko ^a  

^a TAKEDA PHARMACEUTICAL COMPANY LIMITED   (Japan)

Author keywords

Fusion protein; Prolactin releasing peptide; S cyanylation

Indexed keywords

AMMONIA; ARACHIDONIC ACID; BASIC FIBROBLAST GROWTH FACTOR; CYSTEINE; HYBRID PROTEIN; PROLACTIN RELEASING FACTOR; RECOMBINANT DNA; SYNTHETIC PEPTIDE;

AFFINITY CHROMATOGRAPHY; AMIDATION; ANIMAL CELL; ANIMAL TISSUE; ARTICLE; BIOASSAY; CATTLE; CHO CELL; CONTROLLED STUDY; DRUG PURIFICATION; DRUG RECEPTOR BINDING; DRUG SYNTHESIS; GEL FILTRATION CHROMATOGRAPHY; HYPOTHALAMUS; ION EXCHANGE CHROMATOGRAPHY; NONHUMAN; PEPTIDE SYNTHESIS; PROLACTIN RELEASE; RECOMBINANT DNA TECHNOLOGY;

EID: 0031714198     PISSN: 00092363     EISSN: None     Source Type: Journal    
DOI: 10.1248/cpb.46.1490     Document Type: Article

References (17)

1. Hinuma S., Habata Y., Fujii R., Kawamata Y., Hosoya M., Fukusumi S., Kitada C., Masuo Y., Asano T., Sekiguchi M., Matsumoto H., Kurokawa T., Nishimura O., Onda H., Fujino M., Nature (London), 393, 272-276 (1998).
2. Udaka S., Yamagata H., Methods Enzymol., 217, 23-33 (1993).
3. Becker G. W., Hsiung H. M., FEBS Lett., 204 (1), 145-150 (1993).
4. Phillips T. A., Van Bogelen R. A., Neidhardt F. C., J. Bacteriol., 159, 283-287 (1984).
5. Pohlner J., Kramer J., Meyer T. F., Gene, 130 (1), 121-126 (1993).
6. Smith D. B., Johnson K. S., Gene, 67 (1), 31-40 (1988).
7. Seno M., Sasada R., Iwane M., Sudo K., Kurokawa T., Itoh K., Igarashi K., Biochem. Biophys. Res. Commun., 151 (2), 701-708 (1988).
8. Koyama N., Kuriyama M., Amano N., Nishimura O., J. Biotechnol., 32, 273-281 (1994).
9. Nakagawa S., Tamakashi Y., Hamana T., Kawase M., Taketomi S., Ishibashi Y., Nishimura O., Fukuda T., J. Am. Chem. Soc., 116, 5513-5514 (1994).
10. Widmer F., Breddam K., Johansen J. T., Carlsberg Res. Commun., 46, 97-106 (1981).
11. Henriksen D. B., Breddam K., Miller J., Buchardt O., J. Am. Chem. Soc., 114, 1876-1877 (1992).
12. Ray M. V. L., Duyne P. V., Bertelsen A. H., Jackson-Matthews D. E., Sturmer A. M., Merkler D. J., Consalvo A. P., Young S. D., Gilligan J. P., Shields P. P., Biotechnology, 11 (1), 64-70 (1993).
13. Six synthetic oligonucleotides designed to have E. coli bias codons for the bPrRP structural gene and cleavage site were annealed and inserted into Xbal-Aval cloning site of pTB960-7.
14. Watanabe T., Seno M., Sasada R., Igarashi K., Mol. Endocrinol., 4, 869-879 (1990).
15. The cell-free extract was applied to an AF-Heparin Toyopearl 650M column (Tosoh, 3×50cm) equilibrated with 50 mM phosphate buffer (pH 6.0). The column was washed and proteins were eluted with a linear gradient of NaCl (0-2 M).
16. 6M urea was dissolved in the fusion protein solution followed by addition of 0.1M acetic acid. After the addition of 2.4mM of DMAP-CN, the reaction mixture was incubated at 25 °C for 15min. This mixture was applied to a Sephadex G-25 column (Amersham Pharmacia Biotech, 4.6×60cm) equilibrated with 50mM phosphate buffer (pH 6.0). The main fraction was concentrated and 6 M urea and 3 M ammonia were added followed by incubation at 25 °C for 15 min. The mixture was gel filtrated with a Sephadex G-25 column as described above.
17. The reaction mixture was applied to a TSK Gel SP-5PW column (Tosoh, 2.15×15cm) equilibrated with 50mM phosphate buffer (pH 6.0) containing 3M urea. The protein was eluted with a linear gradient of NaCl (0-0.35M). The desired fraction was applied to a C4P-50 column (Showdex, 2.15×30cm) and eluted with a linear gradient of 16-32% acetonitrile in the presence of 0.1% trifluoroacetic acid.