Interaction of Copper(II) Ion with SALI3-2

doi:10.7503/cjcu20120184

Abstract

Abstract:

The effect of CuCl₂ on the growth of the transgenic Arabidopsis carrying Sali3-2 gene was studied on 1/2 MS agar plates. The over-expression of Sali3-2 was elevated the tolerance of transgenic Arabidopsis seedling to Cu²⁺ stress. Sali3-2 gene was then cloned and the SALI3-2 protein deleted signal peptide was expressed and purified. SALI3-2 protein interaction with Cu²⁺ was further studied by immobilized metal ion affinity chromatography(IMAC). The results show that the protein can bind with Cu²⁺. The mechanism of Cu²⁺-SALI3-2 bonding was further analyzed by fuorescence spectrum and circular dichroism(CD) spectrum. Fluorescence analysis shows intrinsic fluorescence of SALI3-2 is quenched with the addition of Cu²⁺, which belongs to static fluorescence quenching. Relatively high affinity of SALI3-2 binding Cu²⁺is revealed with a constant of 8.89×10⁶ and a number of binding sites of 1.6. The secondary structure of SALI3-2 is not changed obviously with the addition of Cu²⁺ as shown in CD spectra.These results suggest that SALI3-2 is a Cu²⁺-binding protein. It may function by altering its conformation through binding Cu²⁺ to trigger the regulation mechanism and elevate the tolerance of cells to Cu²⁺ stress. The research provides the direct insight into molecular mechanism of the role of SALI3-2 in plant tolerance to Cu²⁺stress.

Key words: Cu²⁺, SALI3-2 protein, Metal ion affinity chromatography, Fluorescence spectrum, Circular dichroism spectrum

CLC Number:

Q946
O641.3

TrendMD:

TANG Yu-Lin, GAO Zhan, XU Hong, HE Jian-Zhi, DONG Yi-Han, ZHENG Yi-Zhi. Interaction of Copper(II) Ion with SALI3-2[J]. Chem. J. Chinese Universities, 2013, 34(1): 128.

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Interaction of Copper(II) Ion with SALI3-2

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