A high affinity binding site for [3H]-Dofetilide on human leukocytes.
Certain Class III anti-arrhythmic agents have been shown to interact with human leukocytes and after antigenic and mitogenic activation. We hypothesized that a binding site for the Class III anti-arrhythmic agent, dofetilide, would exist on human leukocytes. Analysis of binding isotherms defined the presence of a single high affinity binding site on mononuclear cells and neutrophils: Kd 26+/-4 nm, Bmax 61+/-14 fmol/10( 6) cells and Kd 33+/-14 nm, Bmax 163+/-45 fmol/10(6) cells, respectively. Other Class III drugs inhibited [3H]-dofetilide binding at physiologically relevant concentrations, but the IC50 values of E4031 and quinidine were significantly higher for leukocytes than for cardiac myocytes. Interestingly, verapamil inhibited [3H]-dofetilide binding to leukocytes, but not to cardiac myocytes at physiologic concentrations (10 microM). Charybdotoxin and tetraethlyammonium inhibited [3H]-dofetilide binding to leukocytes at microM mm concentrations, respectively, however, apamin did not inhibit binding even at 1 microM concentrations. These data suggest that a Ca2+-activated K+ channel, like K(Ca) mini (apamin-insensitive isoform), is a candidate for the leukocyte [3H]-dofetilide binding site. To assess the functional significance of defetilide binding to leukocyte biology, we evaluated fMLP-stimulated superoxide production in the presence or absence of dofetilide. Dofetilide, at 30 nm suppressed of superoxide production. In conclusion, dofetilide binds to human leukocytes at physiologic concentrations and this binding alters leukocyte function possibly through interaction with a Ca2+-activated K+ channel.
The Cardiovascular Research Group, The Universiy of Calgary, 3330 Hospital Drive N.W., Calgary, Alberta, T2N 4N1, Canada.
R
R
Geonzon
D V
DV
Exner
R C
RC
Woodman
L
L
Wang
Z P
ZP
Feng
H J
HJ
Duff
D000818:Animals; D000889:Anti-Arrhythmia Agents / Q000378:metabolism*; D001665:Binding Sites; D018999:Charybdotoxin / Q000494:pharmacology; D006168:Guinea Pigs; D006801:Humans; D007700:Kinetics; D007962:Leukocytes / Q000187:drug effects / Q000378:metabolism*; D009479:Neuropeptides / Q000187:drug effects / Q000378:metabolism; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D010627:Phenethylamines / Q000378:metabolism*; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated; D051670:Shaw Potassium Channels; D013449:Sulfonamides / Q000378:metabolism*; D019789:Tetraethylammonium / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D000889:Anti-Arrhythmia Agents; D009479:Neuropeptides; D010627:Phenethylamines; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D051670:Shaw Potassium Channels; D013449:Sulfonamides; D018999:Charybdotoxin; D019789:Tetraethylammonium; D014700:Verapamil; C063533:dofetilide