title: Bromoenol lactone inhibits voltage-gated Ca2+ and transient receptor potential canonical channels.
abstract: Circulating hormones stimulate the phospholipase Cβ (PLC)/Ca(2+) influx pathway to regulate numerous cell functions, including vascular tone. It was proposed previously that Ca(2+)-independent phospholipase A(2) (iPLA(2))-dependent store-operated Ca(2+) influx channels mediate hormone-induced contractions in isolated arteries, because bromoenol lactone (BEL), a potent irreversible inhibitor of iPLA(2), inhibited such contractions. However, the effects of BEL on other channels implicated in mediating hormone-induced vessel contractions, specifically voltage-gated Ca(2+) (Ca(V)1.2) and transient receptor potential canonical (TRPC) channels, have not been defined clearly. Using isometric tension measurements, we found that thapsigargin-induced contractions were ∼34% of those evoked by phenylephrine or KCl. BEL completely inhibited not only thapsigargin- but also phenylephrine- and KCl-induced ring contractions, suggesting that Ca(V)1.2 and receptor-operated TRPC channels also may be sensitive to BEL. Therefore, we investigated the effects of BEL on heterologously expressed Ca(V)1.2 and TRPC channels in human embryonic kidney cells, a model system that allows probing of individual protein function without interference from other signaling elements of native cells. We found that low micromolar concentrations of BEL inhibited Ca(V)1.2, TRPC5, TRPC6, and heteromeric TRPC1-TRPC5 channels in an iPLA(2)-independent manner. BEL also attenuated PLC activity, suggesting that the compound may inhibit TRPC channel activity in part by interfering with an initial PLC-dependent step required for TRPC channel activation. Conversely, BEL did not affect endogenous voltage-gated K(+) channels in human embryonic kidney cells. Our findings support the hypothesis that iPLA(2)-dependent store-operated Ca(2+) influx channels and iPLA(2)-independent hormone-operated TRPC channels can serve as smooth muscle depolarization triggers to activate Ca(V)1.2 channels and to regulate vascular tone.
authors.affiliation: Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indiana University-Purdue University Indianapolis, 635 Barnhill Dr., MS360A, Indianapolis, IN 46202, USA.
authors.forename: Saikat
authors.initials: S
authors.lastname: Chakraborty
authors.forename: Zachary C
authors.initials: ZC
authors.lastname: Berwick
authors.forename: Paula J
authors.initials: PJ
authors.lastname: Bartlett
authors.forename: Sanjay
authors.initials: S
authors.lastname: Kumar
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Thomas
authors.forename: Michael
authors.initials: M
authors.lastname: Sturek
authors.forename: Johnathan D
authors.initials: JD
authors.lastname: Tune
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Obukhov
mesh_terms: D000818:Animals; D016739:Behavior, Addictive / Q000503:physiopathology; D015220:Calcium Channels / Q000378:metabolism*; D002227:Carbazoles / Q000494:pharmacology; D003042:Cocaine / Q000009:adverse effects / Q000494:pharmacology; D003216:Conditioning, Operant / Q000187:drug effects; D050483:Dopamine Plasma Membrane Transport Proteins / Q000037:antagonists & inhibitors; D018765:Dopamine Uptake Inhibitors / Q000009:adverse effects / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008297:Male; D009281:Naphthalenes / Q000494:pharmacology*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010740:Phospholipases / Q000378:metabolism; D011753:Pyrones / Q000494:pharmacology*; D011869:Radioligand Assay; D051381:Rats; D017207:Rats, Sprague-Dawley; D017480:Receptors, sigma / Q000032:analysis / Q000037:antagonists & inhibitors / Q000187:drug effects; D012646:Self Administration; D013552:Swine; D019284:Thapsigargin / Q000494:pharmacology; D050051:Transient Receptor Potential Channels / Q000378:metabolism*
chemical_list: D015220:Calcium Channels; D002227:Carbazoles; D050483:Dopamine Plasma Membrane Transport Proteins; D018765:Dopamine Uptake Inhibitors; D009281:Naphthalenes; D010726:Phosphodiesterase Inhibitors; D011753:Pyrones; D017480:Receptors, sigma; D050051:Transient Receptor Potential Channels; D019284:Thapsigargin; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; C034931:rimcazole; D010740:Phospholipases; D003042:Cocaine