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Spectral studies of interaction between C-terminal domain on Euplotes Octocarinatus centrin and melittin
Zhao Yaqin 1,Yan Jun 2,Liang Aihua 3,Yang Binsheng 2 *
1.Institute of Molecular Science, Shanxi University
2. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan 030006, China
3.Institute of Molecular Science, Shanxi University, Taiyuan 030006, China
*Correspondence author
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Funding: Natural Science Foundation of Shanxi Province （No.No. 20100110111 and 2011021006-1）, ph.D Programs Foundation of Ministry of Education of the People’s Republic of China （No.No.20091401110007 ）, National Natural Science Foundation of P R China （No.No.20771068 and No. 20901048）
Opened online: 7 November 2011
Accepted by: none
Citation: Zhao Yaqin,Yan Jun,Liang Aihua.Spectral studies of interaction between C-terminal domain on Euplotes Octocarinatus centrin and melittin[OL]. [ 7 November 2011] http://en.paper.edu.cn/en_releasepaper/content/4447906
 
 
Centrin is a low molecular mass (20 kDa) protein that belongs to the EF-hand superfamily of calcium-binding proteins. It plays various roles in a number of different cellular functions. The interactions between C-terminal half of Euplotes octocarinatus centrin (C-EoCen) and melittin (ME) were studied by UV spectra and fluorescence spectra in the presence of 1.0 mM Ca2+. In 100 mM N-2-hydroxyethy-lpiperazine-N-2-ethanesulfonic acid (Hepes) and 150 mM NaCl at pH 7.4, UV absorbance of ME at 280 nm was increased significantly in the presence of Ca2+-saturated C-EoCen (holo-C-EoCen) suggesting the interaction of ME with holo-C-EoCen. In addition, the reaction has been measured by fluorescence spectra under the same experimental conditions. In 100 mM Hepes and 150 mM NaCl at pH 7.4, fluorescence emission of ME appeared at about 353 nm. With the addition of holo-C-EoCen (4.1×10-4 M), fluorescence emission of ME was blue-shifted to 335 nm in virtue of micro-environmental changes of the peptide, indicating that new complex of ME/holo-C-EoCen was formed. Meanwhile, fluorescence emission of ME was increased markedly. On the basis of the result of fluorescence titration curves, the 1:1 stoichiometric ratio of holo-C-EoCen to ME was confirmed. And the conditional binding constant of holo-C-EoCen with ME was calculated to be log KME-holo-C-EoCen = 6.18±0.31.
Keywords:Centrin; Melittin; Fluorescence Spectra; UV spectra
 
 
 

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