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The Study of the Metal-Binding Region in Human Growth Hormone Using Immobilized Metal Ion Affinity Gel-Electrophoresis

Abstract

The zinc(II)-binding affinities of recombinant human growth hormone and two its mutants, 14–33 and 14–95, were studied using Immobilized Metal Ion Affinity Gel-electrophoresis (IMAG). The mutant hormones, composed of polypeptide chain segments of the human and porcine growth hormones, lacked His18, which may be crucial for binding of the intact hormone to the transition metal ions. The mutations did not affect the affinity of human growth hormone to immobilized zinc ions; the structural analysis implied that the human growth hormone contains two IDA–Zn(II) potential sorption sites formed by amino acid residues His21, Asp171, and Glu174 and/or His18 and Glu174.

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REFERENCES

  1. Wells, J.A., Cunnigham, B.C., Fuh, G., Lowman, H.B., Bass, S.H., Mulkerrin, M.G., Ultsch, M., and De Vos, A.M., Recent Prog. Horm. Res., 1993, vol. 48, pp. 253-275.

    Google Scholar 

  2. Cunnigham, B.C., Bass, S., Fuh, G., and Wells, J.A., Science, 1990, vol. 250, pp. 1709-1712.

    Google Scholar 

  3. Somers, W., Ultsch, M., De Vos, A.M., and Kossiakoff, A.A., Nature, 1994, vol. 372, pp. 478-481.

    Google Scholar 

  4. Goubran-Botros, H. and Vijayalakshmi, M.A., Electrophoresis, 1991, vol. 12, pp. 1028-1032.

    Google Scholar 

  5. Baek, W.-O., Haupt, K., Colin, C., and Vijayalakshmi, M.A., Electrophoresis, 1996, vol. 17, pp. 489-492.

    Google Scholar 

  6. Schulga, A.A., Levichkin, I.V., Kurbanov, F.T., Okorokov, A.L., Pozmogova, G.E., and Kirpichnikov, M.P., Nucleic Acids Res., 1994, vol. 22, pp. 3808-3810.

    Google Scholar 

  7. Baek, W.-O. and Vijayalakshmi, M.A., Biochim. Biophys. Acta, 1997, vol. 1336, pp. 394-402.

    Google Scholar 

  8. Anisimova, M.V., Varlamov, V.P., and Vidzhayalakshmi, M.A., Mol. Biol. (Moscow), 1998, vol. 32, pp. 360-361.

    Google Scholar 

  9. Haymore, B.L., Bild, G.S., Salsgiver, W.J., Staten, N.R., and Krivi, G.G., Methods: A Companion to Methods in Enzymology, 1992, vol. 4, pp. 25-40.

    Google Scholar 

  10. Sulkowski, E., Trends Biotechnol., 1989, vol. 53, pp. 1-7.

    Google Scholar 

  11. Sulkowski, E., BioEssays, 1989, vol. 10, pp. 170-175.

    Google Scholar 

  12. Arnold, F.H., Biotechnology, 1991, vol. 9, pp. 151-156.

    Google Scholar 

  13. Jones, T.A., Meth. Enzymol., 1985, vol. 115, pp. 157-171.

    Google Scholar 

  14. Levichkin, I.V., Arsen'eva, D.A., and Shulga, A.A., Mol. Biol. (Moscow), 1998, vol. 32, pp. 317-322.

    Google Scholar 

  15. Skryabin, K.G., Rubtsov, P.M., Gorbulev, V.G., Schulga, A.A., Parsadanian, A.Sh., Kirpichnikov, M.P., Baev, A.A., Pavlovskii, A.G., Borisova, S.N., Vainstein, B.K., and Bulatov, A.A., Protein Structure and Engineering. Series A: Life Sciences, Jardetzky, O., Ed., New York: Plenum, 1990, vol. 183, pp. 309-324.

    Google Scholar 

  16. Bog-Hansen, T.C. and Takeo, K., Electrophoresis, 1980, vol. 1, pp. 67-71.

    Google Scholar 

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Anisimova, M.V., Shulga, A.A., Levichkin, I.V. et al. The Study of the Metal-Binding Region in Human Growth Hormone Using Immobilized Metal Ion Affinity Gel-Electrophoresis. Russian Journal of Bioorganic Chemistry 27, 23–26 (2001). https://doi.org/10.1023/A:1009522917352

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  • DOI: https://doi.org/10.1023/A:1009522917352

  • human growth hormone
  • mutagenesis
  • immobilized metal ion affinity gel-electrophoresis
  • zinc ions