title: Membrane-delimited regulation of novel background K+ channels by MgATP in murine immature B cells.
abstract: In WEHI-231, a representative immature B cell line, Ca(2+) entry is paradoxically augmented by treatment with 2-aminoethoxydiphenyl borate (2-APB), a blocker of inositol 1,4,5-trisphosphate receptor and of nonselective cation channels (Nam, J. H., Yun, S. S., Kim, T. J., Uhm, D.-Y., and Kim, S. J. (2003) FEBS Lett. 535, 113-118). The initial goal of the present study was to elucidate the effects of 2-APB on membrane currents, which revealed the presence of novel K(+) channels in WEHI-231 cells. Under whole-cell patch clamp conditions, 2-APB induced background K(+) current (I(K,bg)) and hyperpolarization in WEHI-231 cells. Lowering of intracellular MgATP also induced the I(K,bg). The I(K,bg) was blocked by micromolar concentrations of quinidine but not by tetraethylammonium. In a single channel study, two types of voltage-independent K(+) channels were found with large (346 picosiemens) and medium conductance (112 picosiemens), named BK(bg) and MK(bg), respectively. The excision of membrane patches (inside-out (i-o) patches) greatly increased the P(o) of BK(bg). In i-o patches, cytoplasmic MgATP (IC(50) = 0.18 mm) decreased the BK(bg) activity, although non-hydrolyzable adenosine 5'-(beta,gamma-imino)triphosphate had no effect. A pretreatment with Al(3+) or wortmannin (50 microm) blocked the inhibitory effects of MgATP. A direct application of phosphoinositide 4,5-bisphosphate (10 microm) inhibited the BK(bg) activity. Meanwhile, the activity of MK(bg) was unaffected by MgATP. In cell-attached conditions, the BK(bg) activity was largely increased by 2-APB. In i-o patches, however, the MgATP-induced inhibition of BK(bg) was weakly reversed by the addition of 2-APB. In summary, WEHI-231 cells express the unique background K(+) channels. The BK(bg)s are inhibited by membrane-delimited elevation of phosphoinositide 4,5-bisphosphate. The activation of BK(bg) would hyperpolarize the membrane, which augments the calcium influx in WEHI-231 cells.
authors.affiliation: Department of Physiology, Center for Molecular Medicine, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Changan-Gu, Cheoncheon-Dong 300, Suwon 440-746, Korea.
authors.forename: Joo Hyun
authors.initials: JH
authors.lastname: Nam
authors.forename: Ji-Eun
authors.initials: JE
authors.lastname: Woo
authors.forename: Dae-Yong
authors.initials: DY
authors.lastname: Uhm
authors.forename: Sung Joon
authors.initials: SJ
authors.lastname: Kim
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000535:Aluminum / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000502:physiology*; D001896:Boron Compounds / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D001896:Boron Compounds; D015221:Potassium Channels; D000255:Adenosine Triphosphate; D000535:Aluminum; C109986:2-aminoethoxydiphenyl borate; D000077191:Wortmannin