Sulfhydryl modulation of K + channels in rat ventricular myocytes

George J Rozanski, Zhi Xu

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Oxidative stress markedly alters protein function through redox modification of sulfhydryl groups present in cysteine residues. To explore the role of redox state in modulating cardiac K + channels, this study examined the effects of sulfhydryl modifiers on the repolarizing transient outward current (I to ) in voltage-clamped myocytes from rat ventricle. Oxidized glutathione (GSSG; 5 mM), an endogenous disulfide that specifically reacts with protein sulfhydryls, decreased maximum I to amplitude from baseline by 49% when added to the external solution (P < 0.05) and by 27% during internal dialysis (P < 0.05). The membrane-impermeable disulfide, 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB) did not alter I to when added to the external solution, but it decreased current amplitude by 31% during internal dialysis (P < 0.05). GSSG-mediated I to inhibition varied in a frequency- and voltage-dependent manner, consistent with a state-dependent blocking mechanism. This phenomenon was also observed in myocytes internally dialyzed with DTNB or Cd 2+ , which also covalently binds to free sulfhydryls. Inhibition of I to by GSSG was not reversed by washout alone, consistent with the stable nature of covalently-modified sulfhydryl groups. However, when myocytes pretreated with GSSG were dialyzed with the reducing agent dithiothreitol, I to amplitude increased significantly by 42% (P < 0.05). These data suggest that α-subunits underlying I to , or associated proteins, have one or more sulfhydryl groups within the cytoplasmic domain that directly modulate channel activity in response to changes in cell redox state. Redox modulation of I to channels may be an important post-translational mechanism contributing to acute changes in cardiac repolarization under conditions of oxidative stress, such as ischemia and reperfusion.

Original languageEnglish (US)
Pages (from-to)1623-1632
Number of pages10
JournalJournal of Molecular and Cellular Cardiology
Volume34
Issue number12
DOIs
StatePublished - Dec 1 2002

Fingerprint

Glutathione Disulfide
Muscle Cells
Oxidation-Reduction
Dithionitrobenzoic Acid
Disulfides
Dialysis
Oxidative Stress
Nitrobenzoates
Proteins
Dithiothreitol
Reducing Agents
Reperfusion
Cysteine
Ischemia
Membranes

Keywords

  • Cardiomyocytes
  • Electrophysiology
  • Patch-clamp
  • Potassium channels
  • Redox
  • Sulfhydryl

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Sulfhydryl modulation of K + channels in rat ventricular myocytes . / Rozanski, George J; Xu, Zhi.

In: Journal of Molecular and Cellular Cardiology, Vol. 34, No. 12, 01.12.2002, p. 1623-1632.

Research output: Contribution to journalArticle

Rozanski, George J ; Xu, Zhi. / Sulfhydryl modulation of K + channels in rat ventricular myocytes In: Journal of Molecular and Cellular Cardiology. 2002 ; Vol. 34, No. 12. pp. 1623-1632.
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