Synthesis of Transmembrane Segment of A Single Spanning Protein, Isk, Forming Potassium Channel and Its Interaction with Phospholipid Bilayr

形成钾通道的单跨膜蛋白 Isk 跨膜片段的合成及其与磷脂双层的相互作用

• 批准号: 05680584
• 负责人: LEE Sannamu
• 金额: $ 1.28万
• 依托单位: Fukuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05680584/
• 关键词: Molecular biology; Ion Channel; Membrane Protein; Peptide synthesis; Potassium Channel; Isk protein; Lipid-protein interaction; 立体構造解析; 生物物理学; 生体膜; ペプチド; 蛋白質-脂質相互作用; 生体膜貫通構造

A membrane protein, Isk, consisting of 130 amino acid residues with only a single transmembrane domain can form a voltage-gated potassium channel which can be opened or closed rapidly by changes in membrane potential in kidney, womb and heart of mammals. The single transmembrane domain is considered to form an alpha-helix by interacting with biomembrane and play a role as a constituent of the pore formed in the assembly of Isk protein monomers within membranes, resulting in the formation of a conducting pore. It is a matter of great interest to make clear the mechanism of the potassium ion selective channel formation by assembling of Isk proteins which have only one transmembrane domain consisting of hydrophobic amino acid residues. Thus, an investigation on the conformation of this portion and behavior of the assembly in membranes will lead to fundamental understanding of the nature of this protein. Therefore we synthesized a peptide of 40 residues containing the transmembrane domain corresponding to residues 38-77 (Isk-O) and its analog which is substituted Ala in Isk-0 for three Gly (Isk-A).In the present study, Isk-0 took beta-structure in neutral and acidic liposomes and showed no channel formation. On the other hand, Isk-A resulted in a gradual increase in conductance followed by a constant conductance but did not exhibit channel-like opening and closing spikes.. Recent publications on the channel forming peptides and proteins have shown that beta-strands seem to be a part of the membrane spanning domain. The membrane spanning Isk domain hypothesized as an alpha-helical structure may adopt beta-structure or a mixture of alpha-helical and beta-structures in biological membranes.Isk-0H-Arg-Asp-Asp-Ser-Lys-Leu-Glu-Ala-Leu-Tyr-Ile-Leu-Met-Val-Leu-Gly-Phe-Phe-Gly-Phe-Phe-Thr-Leu-Gly-Ile-Met-Leu-Ser-Tyr-Ile-Arg-Ser-Lys-Lys-Leu-Glu-His-Ser-His-Asp-0H

Isk是一种由130个氨基酸残基组成的膜蛋白，只有一个跨膜结构域，在哺乳动物的肾脏、子宫和心脏中可形成电压门控钾通道，通过膜电位的变化迅速打开或关闭。单个跨膜结构域被认为通过与生物膜相互作用形成α-螺旋，并作为Isk蛋白单体在膜内组装时形成的孔的组成部分发挥作用，导致形成导电孔。通过组装只有一个由疏水氨基酸残基组成的跨膜结构域的Isk蛋白来阐明钾离子选择性通道的形成机制是一个非常有趣的问题。因此，对该部分的构象和膜中组装行为的研究将导致对该蛋白质性质的基本理解。因此，我们合成了一个40个氨基酸残基的跨膜肽（Isk-0）及其类似物（Isk-0中的三个Gly被Ala取代），Isk-0在中性和酸性脂质体中均为β-结构，无通道形成。另一方面，Isk-A导致电导逐渐增加，随后是恒定的电导，但没有表现出通道样开放和关闭尖峰。最近关于通道形成肽和蛋白质的出版物表明，β链似乎是跨膜结构域的一部分。Isk-0H-Arg-Asp-Asp-Ser-Lys-Leu-Glu-Ala-Leu-Tyr-Ile-Leu-Met-Val-Leu-Gly-Phe-Phe-Gly-Phe-Phe-Thr-Leu-Gly-Ile-Met-Leu-Ser-Tyr-Ile-Arg-Ser-Lys-Lys-Leu-Glu-His-Ser-His-Asp-OH

项目成果

杉原剛介 他4名: "Micokli formation and Camterion binding of mixad sgolom of catiome sunof aetantu haing patluoro carboxylates couterion." Journal of Colloid Interface Science. (in press). (1995)

Kosuke Sugihara 和其他 4 人：“Catiome sunof aetantu haing patluoro carboxylates couterion 的 Micokli 形成和 Camterion 结合”（胶体界面科学杂志）（1995 年）。

李相男他4名: "Two mode ion channels induced by interaction of acidic amphipathic alpha-helical peptides with lipid bilayers" Biochimica et Biophysica Acta. 1191. 181-189 (1994)

Lee Sang-nam 和其他 4 人：“酸性两亲性 α 螺旋肽与脂质双层相互作用诱导的两种模式离子通道”Biochimica et Biophysical Acta 1191. 181-189 (1994)

岩田敏英 他7名: "Design and synthesis if awplipathic 3_<10>-helical paptides and cheir intiraction with phosplielipid bilayers and ion channel formation" Jaurnal of Bislogical Chemistry. 269. 4928-2933 (1994)

Toshihide Iwata 和其他 7 人：“具有磷脂双层和离子通道形成的 awplipathic 3_<10>-螺旋肽的设计和合成”Journal of Bisological Chemistry 269. 4928-2933 (1994)。

李相男他7名: "Effect of salts on conformational change of basic amphipathic peptides from beta-structure to alpha-helix in the presence of phosholipid liposomes and their channel-forming ability" Biochimica et Biophysica Acta. 1151. 76-82 (1993)

Lee Sang-nam 和其他 7 人：“在磷脂脂质体存在下，盐对碱性两亲性肽从 β 结构到 α 螺旋的构象变化及其通道形成能力的影响”Biochimica et Biophysica Acta。 1151。 76-82 (1993)

李相男他4名: "Synthesis of transmembrane segment of a single spanning protein, Isk, forming potassium channel and its interaction with phospholipid bilayer" Peptide Chemistry 1994. 141-144 (1995)

Lee Sang-nam 和其他 4 人：“单个跨越蛋白 Isk 跨膜片段的合成，形成钾通道及其与磷脂双层的相互作用” Peptide Chemistry 1994. 141-144 (1995)