Synthetic Model Peptides for the Study of Structure-Activity Relationship of Ion Channels Using Planar Lipid Bilayer Technique

利用平面脂质双层技术研究离子通道构效关系的合成模型肽

• 批准号: 03671027
• 负责人: ANZAI Kazunori
• 金额: $ 1.34万
• 依托单位: Kyushu 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: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03671027/
• 关键词: ion channel; model peptide; planar bilayer; Amphiphthic; 合成ペプチド; Kチャネル; 平面膜法; 二次構造; 抗菌活性

A model peptide for the H5 region,the putative pore-forming region,of voltage dependent potassium channel(H5 peptide)was synthesized and its interaction with lipid bilayer membranes was investigated by fluorescence spectroscopy,CD measurements,and planar bilayer measurements.The fluorescence of the tryptophan residues present in the peptide was only weakly quenched with KI when the peptide was incorporated into the liposomes.The CD spectra showed that beta-structure was induced in the presence of liposomes.The H5 peptide formed ion channels in planar lipid bilayers.These results support the idea that the H5 region penetrates the membrane and forms pore lining of the voltage dependent potassium channel by taking beta-structure.However,the H5 peptide channel was rather anion selective and its conductance was larger than native potassium channel.These deviations from the native channel may reflect the importance of other membrane spanning regions for the function of the potassium channel.Several model peptides with basic amino acids at every three or four residues of their sequences were synthesized.These peptides are models for S4 segment of voltage dependent sodium channel.These peptides formed cation selective ion channels in planar lipid bilayer membranes.The properties of the channels depended on many parameters such as peptide length,amphipathic properties,the size of the side chains of hydrophobic amino acids,the charge of hydrophilic amino acids,etc.Further quantitative studies for the effect of these parameters on the channel properties will contribute to reveal the interaction of channel proteins with lipid bilayers.

电压依赖性钾离子通道H5区的模型肽合成了H5肽，用荧光光谱、圆二色谱、当肽被掺入脂质体中时，肽中存在的色氨酸残基的荧光仅被KI微弱地猝灭。CD光谱显示β-H5肽在脂质体存在下诱导形成β-结构，H5肽在平面脂质双层中形成离子通道，这些结果支持H5区通过β-结构穿透膜形成电压依赖性钾通道的孔衬的观点，然而，H5肽通道具有较强的阴离子选择性，其电导大于天然钾通道，这与天然钾通道的电导不同本文合成了几种在其序列的每三个或四个残基中含有碱性氨基酸的模型肽，它们是电压依赖性钠通道S4段的模型肽，这些肽在平面脂双层膜中形成阳离子选择性离子通道，通道的性质取决于许多参数，两亲性、疏水性氨基酸侧链的大小、亲水性氨基酸的电荷等。进一步定量研究这些参数对通道性质的影响将有助于揭示通道蛋白与脂双层的相互作用。

项目成果

安西 和紀,桐野 豊: "骨格筋小胞体Caポンプの起電性:新しい実験法による研究の新展開" 実験医学.

Kazunori Anzai、Yutaka Kirino：“骨骼肌内质网钙泵的生电特性：使用新实验方法的研究新进展”实验医学。

Yukio Agawa,Sannamu Lee,Shin Ono,Haruhiko Aoyagi,Motonori Ohno,Takako Taniguchi,Kazunori Anzai,and Yutaka Kirino: "Interaction with phospholipid bilayers,ion channel formation,and antimicrobial activity of basic amphipathic a-herical model peptides of var

Yukio Akawa、Sannamu Lee、Shin Ono、Haruhiko Aoyagi、Motonori Ohno、Takako Taniguchi、Kazunori Anzai 和 Yutaka Kirino：“与磷脂双层的相互作用、离子通道的形成以及 var 的基本两亲性非螺旋模型肽的抗菌活性

T.Iwata,S.Lee,O.Oishi,H.Aoyagi,M.Ohno,K.Shinozaki,K.Anzai,Y.Kirino: "Ion-channel Formation by Various Amphipathic Peptides with the Chain Length to Span Phospholipid Bilayers.Mainly Using 310-Helix Peptide" Peptide Chemistry. 1991. 175-178 (1992)

T.Iwata、S.Lee、O.Oishi、H.Aoyagi、M.Ohno、K.Shinozaki、K.Anzai、Y.Kirino：“各种链长可跨越磷脂双层的两亲性肽形成离子通道。

H.Tokumaru,K.Anzai,T.Abe,Y.Kirino: "Purification of the cardiac 1,4-dihydropyridine receptor using immunoaffinity chromatography with a monodonal antibody agaist the α_2δ subunit of the skeletal muscle DHP receptor" Eur.J.Pharmacol. 227. 363-370 (1992)

H.Tokumaru、K.Anzai、T.Abe、Y.Kirino：“使用针对骨骼肌 DHP 受体 α_2δ 亚基的单克隆抗体进行免疫亲和色谱法纯化心脏 1,4-二氢吡啶受体”Eur.J.Pharmacol . 227. 363-370 (1992)

桐野 豊、安西 和紀: "イオン輸送システム" 蛋白質核酸酵素. 38. (1993)

Yutaka Kirino、Kazunori Anzai：“离子传输系统”蛋白质核酸酶 38。（1993）