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Copper (II) Ions Affect Escherichia coli Membrane Vesicles’ SH-Groups and a Disulfide-Dithiol Interchange Between Membrane Proteins

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Abstract

The SH-groups in Escherichia coli membrane vesicles, prepared from cells grown in fermentation conditions on glucose at slightly alkaline pH, have a role in the F0F1-ATPase operation. The changes in the number of these groups by ATP are observed under certain conditions. In this study, copper ions (Cu2+) in concentration of 0.1 mM were shown to increase the number of SH-groups in 1.5- to 1.6-fold independent from K+ ions, and the suppression of the increased level of SH-groups by ATP was determined for Cu2+ in the presence of K+. Moreover, the increase in the number of SH-groups by Cu2+ was absent as well as the inhibition in ATP-dependent increasing SH-groups number by Cu2+ lacked when vesicles were treated with N-ethylmaleimide (NEM), specific thiol-reagent. Such an effect was not observed with zinc (Zn2+), cobalt (Co2+), or Cu+ ions. The increased level of SH-groups was observed in the hycE or hyfR mutants with defects in hydrogenases 3 or 4, whereas the ATP-dependent increase in the number of these groups was determined in hycE not in hyfR mutants. Both changes in SH-groups number disappeared in the atp or hyc mutants deleted for the F0F1-ATPase or hydrogenase 3 (no activity of hydrogenase 4 was detected in the hyc mutant used). A direct effect of Cu2+ but not Cu+ on the F0F1-ATPase is suggested to lead to conformational changes or damaging consequences, increasing accessible SH-groups number and disturbing disulfide-dithiol interchange within a protein–protein complex, where this ATPase works with K+ uptake system or hydrogenase 4 (Hyd-4); breaks in disulfides are not ruled out.

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Acknowledgment

The study was supported by the grant (#0167) from the Ministry of Education and Science of the Republic of Armenia.

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Authors and Affiliations

  1. Department of Biophysics of the Biological Faculty, Yerevan State University, 1 Alex Manougian Str, Yerevan, 0025, Armenia

    Gayane Kirakosyan, Karen Trchounian, Zaruhi Vardanyan & Armen Trchounian

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  1. Gayane Kirakosyan
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  2. Karen Trchounian
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  3. Zaruhi Vardanyan
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  4. Armen Trchounian
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Correspondence to Armen Trchounian.

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Kirakosyan, G., Trchounian, K., Vardanyan, Z. et al. Copper (II) Ions Affect Escherichia coli Membrane Vesicles’ SH-Groups and a Disulfide-Dithiol Interchange Between Membrane Proteins. Cell Biochem Biophys 51, 45–50 (2008). https://doi.org/10.1007/s12013-008-9014-7

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