Redox-Sensitive Sulfenic Acid Modification Regulates Surface Expression of the Cardiovascular Voltage-Gated Potassium Channel, Kv1.5

Abstract
Rationale: Kv1.5 (KCNA5) is expressed in the heart, where it underlies the IKur current that controls atrial repolarization, and in the pulmonary vasculature, where it regulates vessel contractility in response to changes in oxygen tension. Atrial fibrillation and hypoxic pulmonary hypertension are characterized by down-regulation of Kv1.5 protein expression, as well as with oxidative stress. Formation of sulfenic acid (SOH) on cysteine residues of proteins is an important, dynamic mechanism for protein regulation under oxidative stress. Kv1.5 is widely reported to be redox-sensitive, and the channel possesses six potentially redox-sensitive intracellular cysteines. We therefore hypothesized that sulfenic acid modification of the channel itself may regulate Kv1.5 in response to oxidative stress.
Objective: To investigate how oxidative stress, via redox-sensitive modification of the channel with sulfenic acid, regulates trafficking and expression of Kv1.5.
Methods and Results: Labeling studies with the sulfenic acid–specific probe, DAz, and HRP-streptavidin western blotting demonstrated a global increase in sulfenic acid modified proteins in human patients with atrial fibrillation, as well as sulfenic acid modification to Kv1.5 in the heart. Further studies showed that Kv1.5 is modified with sulfenic acid on a single COOH-terminal cysteine (C581), and the level of sulfenic acid increases in response to oxidant exposure. Using live-cell immunofluorescence and whole-cell voltage clamping, we found that modification of this cysteine is necessary and sufficient to reduce channel surface expression, promote its internalization, and block channel recycling back to the cell surface. Moreover, western blotting demonstrated that sulfenic acid modification is a trigger for channel degradation under prolonged oxidative stress.
Conclusions: Sulfenic acid modification to proteins, which is elevated in diseased human heart, regulates Kv1.5 channel surface expression and stability under oxidative stress, and diverts channel from a recycling pathway to degradation. This provides a molecular mechanism linking oxidative stress and down-regulation of channel expression observed in cardiovascular diseases.
- Atrial fibrillation
- Oxidative stress
- Voltage-gated potassium channels
- Post-translational Modification
- Sulfenic Acid
- Trafficking
- Kv1.5
- Received December 22, 2011.
- Accepted July 27, 2012.
- Copyright © 2012, American Heart Association
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- Redox-Sensitive Sulfenic Acid Modification Regulates Surface Expression of the Cardiovascular Voltage-Gated Potassium Channel, Kv1.5Laurie K. Svoboda, Khalilah G. Reddie, Lian Zhang, Eileen D. Vesely, Elizabeth S. Williams, Sarah M. Schumacher, Ryan P. O'Connell, Robin Shaw, Sharlene M. Day, Justus M. Anumonwo, Kate S. Carroll and Jeffrey R. MartensCirculation Research. 2012;CIRCRESAHA.111.263525, originally published July 27, 2012https://doi.org/10.1161/CIRCRESAHA.111.263525
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- Redox-Sensitive Sulfenic Acid Modification Regulates Surface Expression of the Cardiovascular Voltage-Gated Potassium Channel, Kv1.5Laurie K. Svoboda, Khalilah G. Reddie, Lian Zhang, Eileen D. Vesely, Elizabeth S. Williams, Sarah M. Schumacher, Ryan P. O'Connell, Robin Shaw, Sharlene M. Day, Justus M. Anumonwo, Kate S. Carroll and Jeffrey R. MartensCirculation Research. 2012;CIRCRESAHA.111.263525, originally published July 27, 2012https://doi.org/10.1161/CIRCRESAHA.111.263525