Sigma receptor photolabeling and sigma receptor-mediated modulation of potassium channels in tumor cells

Russell A. Wilke, Rakesh Mehta, Patrick J. Lupardus, Yuenmu Cben, Arnold E. Ruoho, Meyer B. Jackson

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

Recent work has indicated that sigma receptor ligands can modulate potassium channels. However, the only sigma receptor characterized at the molecular level has a novel structure unlike any other receptor known to modulate ion channels. This 26-kDa protein has a hydropathy profile suggestive of a single membrane-spanning domain, with no apparent regions capable of G-protein activation or protein phosphorylation. In the present study patch clamp techniques and photoaffinity labeling were used in DMS-114 cells (a tumor cell line known to express sigma receptors) to investigate the role of the 26-kDa protein in ion channel modulation and probe the mechanism of signal transduction. The sigma receptor ligands N-allylnormetazocine (SKF10047), ditolylguanidine, and (±)-2-(N-phenylethyl-N-propyl)-amino-5- hydroxytetralin all inhibited voltage-activated potassium current (I(K)). Iodoazidococaine (IAC), a high affinity sigma receptor photoprobe, produced a similar inhibition in I(K), and when cell homogenates were illuminated in the presence of IAC, a protein with a molecular mass of 26 kDa was covalently labeled. Photolabeling of this protein by IAC was inhibited by SKF10047 with half-maximal effect at 7 μM. SKF10047 also inhibited I(K) with a similar EC50 (14 μM). Thus, physiological responses to sigma receptor ligands are mediated by a protein with the same molecular weight as the cloned sigma receptor. This indicates that ion channel modulation is indeed mediated by this novel protein. Physiological responses were the same when cells were perfused internally with either guanosine 5'-O-(2-thiodiphosphate) or GTP, indicating that signal transduction is independent of G-proteins. These results demonstrate that ion channels can be modulated by a receptor that does not have seven membrane-spanning domains and does not employ G-proteins. Sigma receptors thus modulate ion channels by a novel transduction mechanism.

Original languageEnglish (US)
Pages (from-to)18387-18392
Number of pages6
JournalJournal of Biological Chemistry
Volume274
Issue number26
DOIs
StatePublished - Jun 25 1999
Externally publishedYes

Fingerprint

sigma Receptors
Potassium Channels
Tumors
Cells
Modulation
Ion Channels
Neoplasms
GTP-Binding Proteins
Proteins
Signal transduction
Ligands
Signal Transduction
Membranes
Phosphorylation
Clamping devices
Molecular mass
Patch-Clamp Techniques
Guanosine Triphosphate
Tumor Cell Line
Labeling

ASJC Scopus subject areas

  • Biochemistry

Cite this

Sigma receptor photolabeling and sigma receptor-mediated modulation of potassium channels in tumor cells. / Wilke, Russell A.; Mehta, Rakesh; Lupardus, Patrick J.; Cben, Yuenmu; Ruoho, Arnold E.; Jackson, Meyer B.

In: Journal of Biological Chemistry, Vol. 274, No. 26, 25.06.1999, p. 18387-18392.

Research output: Contribution to journalArticle

Wilke, Russell A. ; Mehta, Rakesh ; Lupardus, Patrick J. ; Cben, Yuenmu ; Ruoho, Arnold E. ; Jackson, Meyer B. / Sigma receptor photolabeling and sigma receptor-mediated modulation of potassium channels in tumor cells. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 26. pp. 18387-18392.
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