Effect of Clotrimazole on the Pump Cycle of the Na,K-ATPase

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2008
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Bartolommei, Gianluca
Devaux, Nadège
Tadini-Buoninsegni, Francesco
Moncelli, Maria Rosa
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Biophysical Journal. 2008, 95(4), pp. 1813-1825. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1529/biophysj.108.133546
Zusammenfassung

The effect of the antimycotic drug clotrimazole (CLT) on the Na,K-ATPase was investigated using fluorescence and electrical measurements. The results obtained by steady-state fluorescence experiments with the electrochromic styryl dye RH421 were combined with those achieved by a pre-steady-state method based on fast solution exchange on a solid supported membrane that adsorbs the protein. Both techniques are suitable for monitoring the electrogenic steps of the pump cycle and are in general complementary, yielding distinct kinetic information. The experiments show clearly that CLT affects specific partial reactions of the pump cycle of the Na,K-ATPase with an affinity in the low micromolar range and in a reversible manner. All results can be consistently explained by proposing the CLT-promoted formation of an ion-occluded-CLT-bound conformational E2 state, ECLT 2 (Χ2) that acts as a dead-end side track of the pump cycle, where X stands for H1 or K1. Na1 binding, enzyme phosphorylation, and Na1 transport were not affected by CLT, and at high CLT concentrations ~1/3 of the enzyme remained active in the physiological transport mode. The presence of Na1 and K1 destabilized the inactivated form of the Na,K-ATPase.

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570 Biowissenschaften, Biologie
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ISO 690BARTOLOMMEI, Gianluca, Nadège DEVAUX, Francesco TADINI-BUONINSEGNI, Maria Rosa MONCELLI, Hans-Jürgen APELL, 2008. Effect of Clotrimazole on the Pump Cycle of the Na,K-ATPase. In: Biophysical Journal. 2008, 95(4), pp. 1813-1825. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1529/biophysj.108.133546
BibTex
@article{Bartolommei2008Effec-8190,
  year={2008},
  doi={10.1529/biophysj.108.133546},
  title={Effect of Clotrimazole on the Pump Cycle of the Na,K-ATPase},
  number={4},
  volume={95},
  issn={0006-3495},
  journal={Biophysical Journal},
  pages={1813--1825},
  author={Bartolommei, Gianluca and Devaux, Nadège and Tadini-Buoninsegni, Francesco and Moncelli, Maria Rosa and Apell, Hans-Jürgen}
}
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