Sulfhydryl modification inhibits K+ (M) current with kinetics close to acetylcholine in rodent NG108-15 cells

Alla Egorova, Naoto Hoshi, Rimma Knijnik, Mohammad Shahidullah, Minako Hashii, Mami Noda, Haruhiro Higashida

Research output: Contribution to journalArticle

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Abstract

The effects of sulfhydryl reagents on M-type voltage-dependent potassium currents (I(K(M))) were examined in NG108-15 cells transformed to express ml muscarinic acetylcholine receptors (mAChRs), a NGPM1-27 clone. Focal application of glutathione at millimolar concentrations dissolved in acidic solutions caused a transient inward current in NGPM1-27 cells at holding potentials of -30 mV, associated with an inhibition of I(K(M)). The glutathione-induced response was mimicked by cysteine. These effects were also reproduced by superfusion with micromolar concentrations of HgCl2, AgNO3, N-methylmaleimide and p-chloromercuribenzoic acid (pCMB), agents which target protein thiols. Glutathione, HgCl2, AgNO3 and pCMB inhibited the peak conductance of I(K(M)) without shifting the half activating voltage (V(1/2)), which was comparable to the acetylcholine (ACh)-induced response. The voltage dependence of time constants for I(K(M)) deactivation in sulfhydryl reagent-, ACh- and non-treated cells resembled, but differed from that in Ba2+-treated cells. These results reveal that there is an accessible cysteine moiety, but not a disulfide bond, either on the M channel protein itself or on a protein directly involved in agonist-M channel coupling.

Original languageEnglish
Pages (from-to)35-44
Number of pages10
JournalNeuroscience Research
Volume27
Issue number1
DOIs
Publication statusPublished - Mar 25 1997
Externally publishedYes

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Acetylcholine
Rodentia
p-Chloromercuribenzoic Acid
Glutathione
Sulfhydryl Reagents
Mercuric Chloride
Cysteine
Proteins
Muscarinic Receptors
Sulfhydryl Compounds
Disulfides
Potassium
Clone Cells

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

Cite this

Sulfhydryl modification inhibits K+ (M) current with kinetics close to acetylcholine in rodent NG108-15 cells. / Egorova, Alla; Hoshi, Naoto; Knijnik, Rimma; Shahidullah, Mohammad; Hashii, Minako; Noda, Mami; Higashida, Haruhiro.

In: Neuroscience Research, Vol. 27, No. 1, 25.03.1997, p. 35-44.

Research output: Contribution to journalArticle

Egorova, Alla ; Hoshi, Naoto ; Knijnik, Rimma ; Shahidullah, Mohammad ; Hashii, Minako ; Noda, Mami ; Higashida, Haruhiro. / Sulfhydryl modification inhibits K+ (M) current with kinetics close to acetylcholine in rodent NG108-15 cells. In: Neuroscience Research. 1997 ; Vol. 27, No. 1. pp. 35-44.
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