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Thiol-dependent passive K:Cl transport in sheep red blood cells: X. A hydroxylamine-oxidation induced K:Cl flux blocked by diethylpyrocarbonate

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Summary

Hydroxylamine, a potent oxidizing agent used to reverse carbethoxylation of histidine by diethylpyrocarbonate, activated Cl-dependent K flux (K∶Cl cotransport) of low K sheep red blood cells almost sixfold. When K∶Cl cotransport was already stimulated by N-ethylmaleimide, hydroxylamine caused an additional twofold activation suggesting modification of sites different from those thiol alkylated. This conclusion was supported by the finding that hydroxylamine additively augmented also the diamide-induced K∶Cl flux (Lauf, P.K. 1988.J. Membrane Biol. 101:179–188) with dithiothreitol fully reversing the diamide but not the hydroxylamine effect. Stimulation of K∶Cl cotransport by hydroxylamine was completely inhibited by treatment with diethylpyrocarbonate also known to prevent K∶Cl cotransport stimulation by N-ethylmaleimide, both effects being independent of the order of addition. Hence, although the effect of carbethoxy modification on K∶Cl flux cannot be reversed by hydroxylamine and thus excludes histidine as the target for diethylpyrocarbonate, our finding reveals an important chemical determinant of K∶Cl cotransport stimulation by both hydroxylamine oxidation and thiol group alkylation.

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  1. Department of Physiology and Biophysics, Wright State University School of Medicine, 45401-0927, Dayton, Ohio

    P. K. Lauf

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Lauf, P.K. Thiol-dependent passive K:Cl transport in sheep red blood cells: X. A hydroxylamine-oxidation induced K:Cl flux blocked by diethylpyrocarbonate. J. Membrain Biol. 118, 153–159 (1990). https://doi.org/10.1007/BF01868472

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