title: Interaction of propofol with voltage-gated human Kv1.5 channel through specific amino acids within the pore region.
abstract: The intravenous anesthetic propofol affects the function of a diversity of ligand-gated and voltage-gated ion channels. However, there is little information as to whether propofol directly interacts with voltage-gated ion channel proteins to modulate their functions. The Kv1.5 channel is activated by membrane depolarization during action potentials and contributes to atrial repolarization in the human heart. This study was undertaken to examine the effect of propofol on voltage-gated human Kv1.5 (hKv1.5) channel and to elucidate the underlying molecular determinants. Site-directed mutagenesis was carried out through six amino acids that reside within the pore domain of hKv1.5 channel. Whole-cell patch-clamp technique was used to record membrane currents through the wild type and mutant hKv1.5 channels heterologously expressed in Chinese hamster ovary cells. Propofol (≥5 μM) reversibly and concentration-dependently (IC50 of 49.3±9.4 μM; n=6) blocked hKv1.5 current. Propofol-induced block of hKv1.5 current gradually progressed during depolarizing voltage-clamp steps and was enhanced by higher frequency of activation, consistent with a preferential block of the channels in their open state. The degree of current block by propofol was significantly attenuated in T480A, I502A, I508A and V516A, but not in H463C and L510A mutants of hKv1.5 channel. Thus, several amino acids near the selectivity filter (Thr480) or within S6 (Ile502, Ile508 and Val516) are found to be critically involved in the blocking action of propofol. This study provides the first evidence suggesting that direct interaction with specific amino acids underlies the blocking action of propofol on voltage-gated hKv1.5 channel.
authors.affiliation: Department of Anesthesiology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan. Electronic address: akiko77@belle.shiga-med.ac.jp.
authors.forename: Akiko
authors.initials: A
authors.lastname: Kojima
authors.affiliation: Department of Anesthesiology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Department of Physiology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan.
authors.forename: Wei-Guang
authors.initials: WG
authors.lastname: Ding
authors.affiliation: Department of Anesthesiology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan.
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Kitagawa
authors.affiliation: Department of Physiology, Shiga University of Medical Science, Otsu, Shiga 520-2192, Japan. Electronic address: matuurah@belle.shiga-med.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Matsuura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects*; D051666:Kv1.5 Potassium Channel / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers / Q000378:metabolism / Q000494:pharmacology*; D015742:Propofol / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding; D013997:Time Factors; D014162:Transfection
chemical_list: C496981:KCNA5 protein, human; D051666:Kv1.5 Potassium Channel; D026902:Potassium Channel Blockers; D015742:Propofol
keywords: Amino acid; Kv1.5; Mutagenesis; Open channel block; Pore region; Propofol