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Kinetics of luminal proton binding to the SR Ca-ATPase

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2011

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Biophysical Journal. 2011, 101(8), pp. 1896-1904. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1016/j.bpj.2011.09.014

Zusammenfassung

An open membrane preparation containing SR Ca-ATPase was prepared from sarcoplasmic-reticulum vesicles to study the ion binding kinetics in the P-E2 conformation. Because Ca2+ and H+ binding are electrogenic reactions, fluorescent styryl dyes could be used to determine changes in the binding site occupation in equilibrium titration experiments and time-resolved relaxation processes triggered by a pH jump. By photo release from caged proton the pH of the electrolyte could be decreased in a step of 0.1 pH units by a single ultraviolet-laser flash. Analysis of the pH-jump induced relaxation process in the P-E2 conformation showed that three Ca-ATPase-specific processes could be identified, fast H+ binding (τ < 100 μs) and pH-insensitive conformational relaxations after the release of the Ca2+ ion (τ ∼160 ms), and a slow process (τ ∼3.4 s) whose origin could not be unambiguously revealed. The Ca2+-binding affinity in the P-E2 conformation was reduced with increasing pH, a behavior that can be explained by a reversible transition of the empty P-E2 state to an inactivated state of the ion pump. All findings are interpreted in the framework of the Post-Albers pump cycle introduced previously, supplemented by an additional transition to an inhibited state of the ion pump.

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570 Biowissenschaften, Biologie

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ISO 690FIBICH, Andreas, Hans-Jürgen APELL, 2011. Kinetics of luminal proton binding to the SR Ca-ATPase. In: Biophysical Journal. 2011, 101(8), pp. 1896-1904. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1016/j.bpj.2011.09.014
BibTex
@article{Fibich2011-10-19Kinet-16381,
  year={2011},
  doi={10.1016/j.bpj.2011.09.014},
  title={Kinetics of luminal proton binding to the SR Ca-ATPase},
  number={8},
  volume={101},
  issn={0006-3495},
  journal={Biophysical Journal},
  pages={1896--1904},
  author={Fibich, Andreas and Apell, Hans-Jürgen}
}
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