Donghee Kim

TBAK-1 and TASK-1, two-pore K(+) channel subunits: kinetic properties and expression in rat heart.

A mammalian K+ channel subunit (TBAK-1/TASK-1) containing two pore domains and four transmembrane segments and whose mRNA is highly expressed in the heart has been cloned recently. TBAK-1 and TASK-1 are identical except for the additional nine amino acids in the NH2 terminus of TBAK-1. We examined their kinetic properties, pH sensitivity, and regional cardiac mRNA expression and determined whether a native cardiac K+ channel with similar kinetic properties was present. When TBAK-1 or TASK-1 was transiently expressed in COS-7 cells, time- and voltage-independent whole cell currents were observed. Single-channel conductances of TBAK-1 and TASK-1 were 14.6 ± 1.0 and 13.8 ± 2.8 pS, respectively, at -80 mV in 140 mM extracellular K+, and the mean open times were 0.8 ± 0.1 and 0.6 ± 0.1 ms, respectively. Both TBAK-1 and TASK-1 were highly sensitive to extracellular pH such that a decrease from 7.2 to 6.4 reduced their open probability ( P o) by 81 ± 14% and 80 ± 16%, whereas a decrease in intracellular pH from 7.2 to 6.4 reduced the P o by 42 ± 10% and 47 ± 12%, respectively. TBAK-1/TASK-1 mRNA was expressed in all regions of the rat heart, with the highest level of expression in the right atrium. A 14-pS K+channel with kinetic properties similar to those of TBAK-1/TASK-1 was identified in rat atrial and ventricular cells. These results indicate that TBAK-1/TASK-1 represents a functional native K+ channel in the rat heart.

Synthesis of a novel series of 2-alkylthio substituted naphthoquinones as potent acyl-CoA: Cholesterol acyltransferase (ACAT) inhibitors

We report a new series of naphthoquinone derivatives as potent ACAT inhibitors, which were obtained through structural variations of previously disclosed lead 1. Several analogs represented by 3i-l, 4k-m, 6a-n, 7a, and 7i demonstrated potent human macrophage ACAT inhibitory activity by a cell-based reporter assay with human HepG2 cell lines. In particular, compounds 4l and 6j emerged as highly potent inhibitors, exhibiting significantly high inhibitory potencies with IC50 values of 0.44 μM and 0.6 μM, respectively. Moreover, compound 4l significantly reduced the accumulation of cellular cholesterol in HepG2 cell lines.

Inhibition of ATP-induced increase in muscarinic K+ current by trypsin, alkaline pH, and anions

In atrial cells, the open probability of G protein-activated ACh-sensitive K+ (KACh) channels can be increased approximately fivefold by intracellular ATP (ATPi). Using inside-out patches, we examined how proteases, changes in intracellular pH, and different anions affect G protein-mediated activation and ATP-induced stimulation of the KACh channel. Treatment with trypsin (0.5 mg/ml) removed the GTP dependence of the KACh channel and abolished the ATP-induced stimulation. Intracellular GTP activated KACh channels at all intracellular pH values tested (6.0-8.0), with the concentration at which half-maximal activation (K1/2) occurred ranging from 0.3 (pH 8.0) to 6.7 (pH 6.0) microM. However, the ATPi-induced increase in KACh channel activity was inhibited at pH 8. 0 (K1/2 = pH 7.4). All anions tested except sulfate, phosphate, fluoride, and iodide supported GTP-induced activation. Of the anions that supported GTP-induced activation, only citrate blocked the ATP-induced stimulation of the KACh channel. These results indicate that the GTP- and ATP-mediated effects on the KACh channel use separate signaling pathways. The ATP-mediated effect involves a trypsin- and pH-sensitive mechanism.