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Point amino acid substitutions in the Ca2+-binding sites of recoverin: II. The unusual behavior of the protein upon the binding of calcium ions


The structural properties of myristoylated forms of recombinant recoverin of the wild type and of its mutants with damaged second and/or third Ca2+-binding sites were studied by fluorimetry and circular dichroism. The interaction of wild-type recoverin with calcium ions was shown to induce unusual structural rearrangements in its molecule. In particular, protein binding with Ca2+ ions results in an increase in the mobility of the environment of Trp residues, in hydrophobicity, and in thermal stability (its thermal transition shifts by 15°C to higher temperatures) but has almost no effect on its secondary structure. Similar structural changes induced by Ca2+ are also characteristic of the -EF2 mutant of recoverin whose second Ca2+-binding site is modified and cannot bind calcium ions. The structural properties of the -EF3 and -EF2,3 mutants (whose third or simultaneously second and third Ca2+-binding sites, respectively, are modified and damaged) are practically indifferent to the presence of calcium ions.

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8-anilino-1-naphthalenesulfonic acid


circular dichroism


ethylene glycol-bis(β-aminoethyl ether)-N,N,N′N′-tetraacetic acid


recombinant recoverin of wild type


  1. Permyakov, S.E., Senin, I.I., Uverskii, V.N., Cherskaya, A.M., Shul’ga-Morskoy, S.V., Zinchenko, D.V., Alekseev, A.M., Zargarov, A.A., Lipkin, V.M., Filippov, P.P., and Permyakov, E.A.,Bioorg. Khim., 1999, vol. 25, pp. 742–746.

    CAS  Google Scholar 

  2. Permyakov, E.A., Yarmolenko, V.V., Emelyanenko, V.I., Burstein, E.A., Gerday, C., and Closset, J.,Eur. J. Biochem., 1980, vol. 109, pp. 307–315.

    PubMed  Article  CAS  Google Scholar 

  3. Permyakov, E.A., Yarmolenko, V.V., Kalinichenko, L.P., Morozova, L.A., and Burstein, E.A.,Biochem. Biophys. Res. Commun., 1981, vol. 100, pp. 191–197.

    PubMed  Article  CAS  Google Scholar 

  4. Ames, J.B., Ishima, R., Tanaka, T., Gordon, J.I., Stryer, L., and Ikura, M.,Nature, 1997, vol. 389, pp. 198–202.

    PubMed  Article  CAS  Google Scholar 

  5. Tanaka, T., Ames, J.B., Harvey, T.S., Stryer, L., and Ikura, M.,Nature, 1995, vol. 376, p. 444.

    PubMed  Article  CAS  Google Scholar 

  6. Hughes, R.E., Brzovic, P.S., Klevit, R.E., and Hurley, J.B.,Biochemistry, 1995, vol. 34, pp. 11 410–11 416.

    Article  CAS  Google Scholar 

  7. Kataoka, M., Mihara, K., and Tokunaga, F.,J. Biochem., 1993, vol. 114, pp. 535–540.

    PubMed  CAS  Google Scholar 

  8. Stryer, L.,J. Mol. Biol., 1965, vol. 13, pp. 482–495.

    PubMed  CAS  Article  Google Scholar 

  9. Semisotnov, G.V., Rodionova, N.V., Razgulyaev, O.I., Uversky, V.N., Gripas, A.F., and Gilmanshin, R.I.,Biopolymers, 1991, vol. 31, pp. 119–128.

    PubMed  Article  CAS  Google Scholar 

  10. Permyakov, E.A.,Kal’tsiisvyazyvayushchie belki (Calcium-Binding Proteins), Moscow: Nauka, 1993.

    Google Scholar 

  11. Permyakov, E.A., Shnyrov, V.L., Kalinichenko, L.P., Kuchar, A., Reyzer, I.L., and Berliner, L.J.,J. Prot. Chem., 1991, vol. 10, pp. 577–584.

    Article  CAS  Google Scholar 

  12. Leontiev, V.V., Uversky, V.N., Permyakov, E.A., and Murzin, A.G.,Biochim. Biophys. Acta, 1993, vol. 1162, pp. 84–88.

    PubMed  CAS  Google Scholar 

  13. Garnier, C., Protasevich, I., Gilli, R., Tsvetkov, F., Lobachov, V., Peyrot, V., Briand, C., and Makarov, A.,Biochem. Biophys. Res. Commun., 1998, vol. 249, pp. 197–201.

    PubMed  Article  CAS  Google Scholar 

  14. Permyakov, E.A. and Burstein, E.A.,Biophys. Chem., 1984, vol. 19, pp. 265–271.

    PubMed  Article  CAS  Google Scholar 

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Correspondence to E. A. Permyakov.

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For the communication I, see [1].

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Uversky, V.N., Permyakov, S.E., Senin, I.I. et al. Point amino acid substitutions in the Ca2+-binding sites of recoverin: II. The unusual behavior of the protein upon the binding of calcium ions. Russ J Bioorg Chem 26, 152–156 (2000).

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Key words

  • photoreception
  • calcium-binding proteins
  • recoverin
  • site-directed mutagenesis
  • fluorescence
  • circular dichroism