한국가축번식학회지 (Korean Journal of Animal Reproduction)

한국동물번식학회 (The Korean Society of Animal Reproduction)
권호 표지

COOH-Terminal Animo Acids of Tethered-Buman Glycoprotein Bormone $\alpha$-Subunit Play an Important Role for Secretion

  • Min, K.S (Graduate School of Bio & Information Technology, Hankyong national University) ;
  • Yoon, J.K. (Dept. of Animal Life and Resource, Hanhyong National University)
  • 발행 : 2002.12.01
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초록

Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH. hCG TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. To determine u and $\beta$ -subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCC and -FSH molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed. To determine the importance of $\alpha$ COOH -terminal amino acid, we also deleted the $\alpha$ COOH-terminal amino acids. The expressing vectors were transfected into CHO-K 1 cells. The tethered-wthCG and -wtFSH was efficiently secreted. The $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants had no secretion. These results are the first conclusive evidence that COOH-terminal amino acids are very important for secretion in human glycoprotein hormone $\alpha$-subunit. These results demonstrated that the $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants could be play a pivotal role in the secretion of tethered-molecule.

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참고문헌

  1. Ben-Menahem, D., Jablonka-Shariff, A., Hyde, R. K., Pixley, M. R., Srivastava, S., Berger, P. and Boime, I. 2001. The position of the ${\alpha}$ and ${\beta}$ subunits in a single chain variant of human chorionic gonadotropin affects the heterodimeric interaction of the subunits and receptor -binding epitopes. J. BioI. Chem., 276:29871-29879
  2. Chen, F., Wang, Y. and Puett, D. 1992. The carboxy-terminal region of the glycoprotein hormone alpha-subunit: contributions to receptor binding and signaling in human chorionic gonadotropin. Mol. Endocrinol., 6:914-919 https://doi.org/10.1210/me.6.6.914
  3. Matzuk, M. M. and Boime, I. 1988. The role of asparagines-linked oligosaccharides of the a -subunit in the secretion and assembly of human chorionic gonadotropin. J. Cell Biol., 106:1049-1059 https://doi.org/10.1083/jcb.106.4.1049
  4. Fiddes, J. and Goodman, H. M. 1979. Isolation, cloning and sequence analysis of the cDNA for the a-subunit of human chorionic gonadotropin. Nature, 281:351-356 https://doi.org/10.1038/281351a0
  5. Min, K. S. 2002. On the biological functions of equine chorionic gonadotropin. Korean J. Animal Reprod., 26:299-308
  6. Min, K. S. 2000. On the secretion and functions of equine chorionic gonadotropin. Korean J. Animal Reprod., 24:125-142
  7. Min, K. S. 2001. Biosynthesis of a biological active single chain equine chorionic gonadotropin. J. Life. Science, 11: 103-107
  8. Min, K. S., Hattori, N., Aikawa, J. I., Shiota, K. and Ogawa. T. 1996. Site-directed mutagenesis of recombinant equine chorionic gonadotropinl luteinizing hormone: differential role of oligosaccharides in luteinizing hormone- and follicle -stimulating hormone-like activities. Endocrine J., 43:585-593 https://doi.org/10.1507/endocrj.43.585
  9. Min, K. S., Kang, M. H., Yoo, J. T., Seong, H. H., Jin, H. J. and Chang, W. K. 2003. Production of biological active single chain bovine LH and FSH. Asian-Aus. J. Animal Sci., 4:498 -503
  10. Min, K. S., Shiota, K., Saneyoshi, T., Hirosawa, M. and Ogawa, T. 1997. Differential role of oligosacchairdes in equine chorionic gonadotropin( eCG)!luteinizing hormone(LH) to express follicle stimulating hormone(FSH)-like and LH -like activities. J. Reprod. & Dev., 43:177-179
  11. Saneyoshi, T., Min, K. S., Ma, X. J., Nambo, Y., Hiyama, T., Tanaka, T. and Shiota, K. 2001. Equine follicle-stimulating hormone: Molecular cloning of .B-subunit and biological role of the asparagines-linked oligosaccharide at asparagine.' of a-subunit. BioI. Reprod., 65: 1686-1690
  12. Sugahara, T., Grootenhuis, P. D. J., Sato, A., Kudo, M., Ben-Menahem, D., Pixley, M. R., Hsueh, A. 1. W. and Boime, I. 1996. Expression of biologically active fusion genes encoding the common a subunit and either the CG .B or FSH.B subunit: role of a linker sequence. Mol. Cell. Endocrinol., 125:71-77
  13. Sugahara, T., Pixley, M. R., Minami, S., Perlas, E., Ben-Menahem, D., Hsueh, A. J. W. and Boime, I. 1995. Biosynthesis of a biologically active single peptide chain containing the human common a and chorionic gonadotropin .B subunits in tandem. Proc. Natl. Acad. Sci., 92:2041-2045 https://doi.org/10.1073/pnas.92.6.2041
  14. Yoo, J. K., Ji, 1. H. and Ji, T. H. 1991. Conversion of Lysine 91 to methionine of Glutamic acid in human choriogonadotropin a results in the loss of camp inducibility. J. Biol. Chem., 266:17741-17743
  15. Yoo, J. K., Zeng, H., Ji, I. H., Murdoch, W. J. and Ji, T. H. 1993. COOH-terminal amino acids of the a subunit play common and different roles in human choriogonadotropin and follitropin. J. BioI. Chem., 268:13034-13042