Ra. Wilke et al., Sigma receptor photolabeling and sigma receptor-mediated modulation of potassium channels in tumor cells, J BIOL CHEM, 274(26), 1999, pp. 18387-18392
Recent work has indicated that sigma receptor ligands can modulate potassiu
m channels. However, the only sigma receptor characterized at the molecular
level has a novel structure unlike any other receptor known to modulate io
n channels. This 26-kDa protein has a hydropathy profile suggestive of a si
ngle membrane-spanning domain, with no apparent regions capable of G-protei
n activation or protein phosphorylation, In the present study patch clamp t
echniques and photoaffinity labeling were used in DMS-114 cells (a tumor ce
ll 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 tr
ansduction, The sigma receptor Ligands N-allylnormetazocine (SKF10047), dit
olyl-guanidine, and (+/-)-2-(N-phenylethyl-N-propyl)-amino-5-hy- droxytetra
lin all inhibited voltage-activated potassium current (I-K). Iodoazidococai
ne (IAC), a high affinity sigma receptor photoprobe, produced a similar inh
ibition 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. P
hotolabeling of this protein by IAC was inhibited by SKF10047 with half-max
imal effect at 7 mu M. SKF10047 also inhibited I-K with a similar EC50 (14
mu M). 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 in
ternally with either guanosine 5'-O-(2-thiodiphosphate) or GTP, indicating
that signal transduction is independent of G-proteins, These results demons
trate that ion channels can be modulated by a receptor that does not have s
even membrane-spanning domains and does not employ G-proteins, Sigma recept
ors thus modulate ion channels by a novel transduction mechanism.