Oxidative regulation of large conductance calcium-activated potassium channels.
Reactive oxygen/nitrogen species are readily generated in vivo, playing roles in many physiological and pathological conditions, such as Alzheimer's disease and Parkinson's disease, by oxidatively modifying various proteins. Previous studies indicate that large conductance Ca(2+)-activated K(+) channels (BK(Ca) or Slo) are subject to redox regulation. However, conflicting results exist whether oxidation increases or decreases the channel activity. We used chloramine-T, which preferentially oxidizes methionine, to examine the functional consequences of methionine oxidation in the cloned human Slo (hSlo) channel expressed in mammalian cells. In the virtual absence of Ca(2+), the oxidant shifted the steady-state macroscopic conductance to a more negative direction and slowed deactivation. The results obtained suggest that oxidation enhances specific voltage-dependent opening transitions and slows the rate-limiting closing transition. Enhancement of the hSlo activity was partially reversed by the enzyme peptide methionine sulfoxide reductase, suggesting that the upregulation is mediated by methionine oxidation. In contrast, hydrogen peroxide and cysteine-specific reagents, DTNB, MTSEA, and PCMB, decreased the channel activity. Chloramine-T was much less effective when concurrently applied with the K(+) channel blocker TEA, which is consistent with the possibility that the target methionine lies within the channel pore. Regulation of the Slo channel by methionine oxidation may represent an important link between cellular electrical excitability and metabolism.
Department of Physiology and Biophysics, The University of Iowa, Iowa City, Iowa 52242, USA.
X D
XD
Tang
H
H
Daggett
M
M
Hanner
M L
ML
Garcia
O B
OB
McManus
N
N
Brot
H
H
Weissbach
S H
SH
Heinemann
T
T
Hoshi
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D018929:Cell Culture Techniques; D002700:Chloramines / Q000494:pharmacology; D004594:Electrophysiology; D006801:Humans; D015640:Ion Channel Gating; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008715:Methionine / Q000378:metabolism; D010084:Oxidation-Reduction; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000378:metabolism / Q000502:physiology*; D024681:Potassium Channels, Calcium-Activated; D017382:Reactive Oxygen Species; D014105:Tosyl Compounds / Q000494:pharmacology; D015854:Up-Regulation
D002700:Chloramines; D051036:Large-Conductance Calcium-Activated Potassium Channels; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D017382:Reactive Oxygen Species; D014105:Tosyl Compounds; C016300:chloramine-T; D008715:Methionine; D002118:Calcium