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Metal ion-binding properties of wild-type and mutant D37K of ciliate Euplotes octocarinatus centrin

  • Article
  • Inorganic Chemistry
  • Published:
Chinese Science Bulletin

Abstract

Ciliate Euplotes octocarinatus centrin (EoCen) is an EF-hand calcium-binding protein closely related to the prototypical calcium sensor protein calmodulin. The first amino acid of the Ca2+-binding loops found in the EF-hand calcium-binding proteins is a highly conserved aspartic acid residue. The D37K mutant was produced to elucidate the metal binding role of the first aspartic acid of the EF-loop I of EoCen. Aromatic-sensitized Tb3+ fluorescence results indicated that the metal binding ability of loop I was lost due to the D37K mutation. Based on fluorescence titration curves of Lu2-D37K, the conditional binding constants of the EoCen loop II were quantitatively found to be K II = (1.61 ± 0.04) × 105 L mol−1 and K II = (3.52 ± 0.08) × 102 L mol−1 with Tb3+ and Ca2+, respectively. Using 2-p-toluidinylnaphthalene-6-sulfonate as a hydrophobic probe, exposure of the hydrophobic surface upon metal binding was found to be significantly reduced for the metal ion-saturated EoCen D37K mutant.

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Authors and Affiliations

  1. Institute of Molecular Science, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, China

    Wen Liu, Lian Duan, YaQin Zhao & BinSheng Yang

  2. Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030006, China

    AiHua Liang

Authors
  1. Wen Liu
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  2. Lian Duan
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  3. YaQin Zhao
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  4. AiHua Liang
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  5. BinSheng Yang
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Correspondence to BinSheng Yang.

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Liu, W., Duan, L., Zhao, Y. et al. Metal ion-binding properties of wild-type and mutant D37K of ciliate Euplotes octocarinatus centrin. Chin. Sci. Bull. 55, 3118–3122 (2010). https://doi.org/10.1007/s11434-010-3279-0

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  • DOI: https://doi.org/10.1007/s11434-010-3279-0

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