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Russian Journal of Bioorganic Chemistry

, Volume 26, Issue 3, pp 152–156 | Cite as

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

  • V. N. Uversky
  • S. E. Permyakov
  • I. I. Senin
  • A. M. Cherskaya
  • S. V. Shulga-Morskoy
  • D. V. Zinchenko
  • A. M. Alekseev
  • A. A. Zargarov
  • V. M. Lipkin
  • P. P. Philippov
  • E. A. Permyakov
Article
  • 25 Downloads

Abstract

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.

Key words

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

Abbreviations

ANS

8-anilino-1-naphthalenesulfonic acid

CD

circular dichroism

EGTA

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

wt-recoverin

recombinant recoverin of wild type

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Copyright information

© MAIK “Nauka/Interperiodica” 2000

Authors and Affiliations

  • V. N. Uversky
    • 1
  • S. E. Permyakov
    • 1
  • I. I. Senin
    • 2
  • A. M. Cherskaya
    • 1
  • S. V. Shulga-Morskoy
    • 3
  • D. V. Zinchenko
    • 3
  • A. M. Alekseev
    • 3
  • A. A. Zargarov
    • 3
  • V. M. Lipkin
    • 3
  • P. P. Philippov
    • 2
  • E. A. Permyakov
    • 1
  1. 1.Institute for Biological Instrument MakingRussian Academy of SciencesPushchino, Moscow oblastRussia
  2. 2.Belozersky Institute of Physicochemical BiologyMoscow State UniversityVorob’evy gory, MoscowRussia
  3. 3.Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Pushchino BranchRussian Academy of SciencesPushchino, Moscow oblastRussia

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