Zn2+-dependent redox switch in the intracellular T1-T1 interface of a Kv channel.
The thiol-based redox regulation of proteins plays a central role in cellular signaling. Here, we investigated the redox regulation at the Zn(2+) binding site (HX(5)CX(20)CC) in the intracellular T1-T1 inter-subunit interface of a Kv4 channel. This site undergoes conformational changes coupled to voltage-dependent gating, which may be sensitive to oxidative stress. The main results show that internally applied nitric oxide (NO) inhibits channel activity profoundly. This inhibition is reversed by reduced glutathione and suppressed by intracellular Zn(2+), and at least two Zn(2+) site cysteines are required to observe the NO-induced inhibition (Cys-110 from one subunit and Cys-132 from the neighboring subunit). Biochemical evidence suggests strongly that NO induces a disulfide bridge between Cys-110 and Cys-132 in intact cells. Finally, further mutational studies suggest that intra-subunit Zn(2+) coordination involving His-104, Cys-131, and Cys-132 protects against the formation of the inhibitory disulfide bond. We propose that the interfacial T1 Zn(2+) site of Kv4 channels acts as a Zn(2+)-dependent redox switch that may regulate the activity of neuronal and cardiac A-type K(+) currents under physiological and pathological conditions.
Wang
Guangyu
G
Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA. Guang-yu.Wang@jefferson.edu
Strang
Candace
C
Pfaffinger
Paul J
PJ
Covarrubias
Manuel
M
D000818:Animals; D003545:Cysteine / Q000235:genetics / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D017136:Ion Transport / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D020125:Mutation, Missense; D009206:Myocardium / Q000378:metabolism; D009417:Nerve Tissue / Q000378:metabolism; D009569:Nitric Oxide / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D051672:Shal Potassium Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014981:Xenopus; D015032:Zinc / Q000737:chemistry / Q000378:metabolism*
D004220:Disulfides; D051672:Shal Potassium Channels; D009569:Nitric Oxide; D015032:Zinc; D003545:Cysteine