Type of Publication: | Journal article |
Author: | Leitenmaier, Barbara; Witt, Annelie; Witzke, Annabell; Stemke, Anastasia; Meyer-Klaucke, Wolfram; Kroneck, Peter; Küpper, Hendrik |
Year of publication: | 2011 |
Published in: | Biochimica et Biophysica Acta (BBA) - Biomembranes ; 1808 (2011), 10. - pp. 2591-2599. - ISSN 0005-2736 |
Pubmed ID: | 21621506 |
DOI (citable link): | https://dx.doi.org/10.1016/j.bbamem.2011.05.010 |
Summary: |
TcHMA4 (GenBank no. AJ567384), a Cd/Zn transporting ATPase of the P1B-type (=CPx-type) was isolated and purified from roots of the Cd/Zn hyperaccumulator Thlaspi caerulescens. Optimisation of the purification protocol, based on binding of the natural C-terminal His-tag of the protein to a Ni-IDA metal affinity column, yielded pure, active TcHMA4 in quantities sufficient for its biochemical and biophysical characterisation with various techniques. TcHMA4 showed activity with Cu(2+), Zn(2+) and Cd(2+) under various concentrations (tested from 30 nM to 10 μM), and all three metal ions activated the ATPase at a concentration of 0.3 μM. Notably, the enzyme worked best at rather high temperatures, with an activity optimum at 42 °C. Arrhenius plots yielded interesting differences in activation energy. In the presence of zinc it remained constant (EA=38 kJ⋅mol−1) over the whole concentration range while it increased from 17 to 42 kJ⋅mol−1 with rising copper concentration and decreased from 39 to 23 kJ⋅mol−1 with rising cadmium concentration. According to EXAFS the TcHMA4 appeared to bind Cd(2+) mainly by thiolate sulphur from cysteine, and not by imidazole nitrogen from histidine.
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Subject (DDC): | 570 Biosciences, Biology |
Keywords: | Metal specificity, Hyperaccumulator, Natural overexpression, Cd/Zn ATPase, Activation energy, Temperature optimum |
Bibliography of Konstanz: | Yes |
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LEITENMAIER, Barbara, Annelie WITT, Annabell WITZKE, Anastasia STEMKE, Wolfram MEYER-KLAUCKE, Peter KRONECK, Hendrik KÜPPER, 2011. Biochemical and biophysical characterisation yields insights into the mechanism of TcHMA4, a Cd/Zn transporting ATPase purified from the hyperaccumulator plant Thlaspi caerulescens. In: Biochimica et Biophysica Acta (BBA) - Biomembranes. 1808(10), pp. 2591-2599. ISSN 0005-2736. Available under: doi: 10.1016/j.bbamem.2011.05.010
@article{Leitenmaier2011-10Bioch-15520, title={Biochemical and biophysical characterisation yields insights into the mechanism of TcHMA4, a Cd/Zn transporting ATPase purified from the hyperaccumulator plant Thlaspi caerulescens}, year={2011}, doi={10.1016/j.bbamem.2011.05.010}, number={10}, volume={1808}, issn={0005-2736}, journal={Biochimica et Biophysica Acta (BBA) - Biomembranes}, pages={2591--2599}, author={Leitenmaier, Barbara and Witt, Annelie and Witzke, Annabell and Stemke, Anastasia and Meyer-Klaucke, Wolfram and Kroneck, Peter and Küpper, Hendrik} }
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