Synthesis and Structure-Function Relationship Study of Pore Structures emulated Ligand-gated Ionic Channel

配体门控离子通道模拟孔结构的合成及构效关系研究

• 批准号: 07808073
• 负责人: YAMAZAKI Masahito
• 金额: $ 1.41万
• 依托单位: Shizuoka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07808073/
• 关键词: ionic channel protein; design of new proteins; ion permeability; helix bundle structure; single channel recording; phase stability of biomembranes; polymorphism of F-actin; intermembrane interaction of biomembranes; 人工蛋白質; グリシンリセプター; カルシウムATPase

(1)A peptide with the predicted transmembrane sequence of botulinum neurot6xin A was synthesized. Single-channel recording data shows that it forms cation-selective ionic channels in lipid bilayers.(2)A few template-assembled helix bundle proteins emulated the pore structure of acetyicholine receptor were synthesized by a new chemoselective-ligation method developed by us.(3)Ion permeability of a membrane with soft polar interfaces with fixed charges and dipoles were investigated theoretically.(4)A peptide with the predicted transmembrane sequence of M2 proteins of influenza virus A was synthesized, and its structure in bilayer membranes and function of proton transport were studied.(5)Effect of co-ions on proton conductance of gramicidin A channel were investigated by the single channel recording method, and results were analyzed by a theoretical model.(6)Stability and structure of non-bilayer structures of biomembranes such as hexagonal II and interdigitated gel phases were studied by X-ray diffraction and fluorescence spectroscopy. These results indicate that interaction free energy between segments of membrane surface and solvents plays an important role in the stability of these membranes.(7)Polymorphism of F-actin assembly was studied, and its quantitative phase diagram was made.

(1)合成了具有预测的肉毒杆菌神经6xin A跨膜序列的肽。单通道记录数据表明，它在脂质双层中形成阳离子选择性离子通道。(2)采用新的化学选择性连接方法合成了几个模拟乙酰胆碱受体孔结构的模板组装螺旋束蛋白。(3)从理论上研究了具有固定电荷和偶极子的软极性界面膜的离子渗透性。(4)合成了具有预测的甲型流感病毒M2蛋白跨膜序列的肽段，并研究了其在双层膜中的结构和质子转运功能。(5)采用单通道记录法研究了共离子对gramicidin A通道质子电导的影响，并用理论模型对结果进行了分析。(6)利用x射线衍射和荧光光谱技术研究了六方凝胶相和交叉指状凝胶相等生物膜非双层结构的稳定性和结构。这些结果表明，膜表面与溶剂之间的相互作用自由能对膜的稳定性起着重要的作用。(7)研究了F-actin组装的多态性，并绘制了其定量相图。

项目成果

Hatanaka, Y.: "Osmotic stress induces a phasetransition from interdigitated gel phase to bilayer gel phase in multilamellar vesicles of dihexadecylphosphatidylcholine" Biophys.Chem.vol.65. 229-233 (1997)

Hatanaka, Y.：“渗透应力诱导二十六烷基磷脂酰胆碱多层囊泡中从指叉凝胶相到双层凝胶相的相变”Biophys.Chem.vol.65。

Kinoshita,K.: "Organic solvents induce・interdigitated gelstructures in multilamellar vesicles of dipalmitoyl phosphatidyl choline" Biochim.Biophys.Acta. 1284. 233-239 (1996)

Kinoshita, K.：“有机溶剂在二棕榈酰磷脂酰胆碱的多层囊泡中诱导交指状凝胶结构”Biochim.Biophys.Acta. 1284. 233-239 (1996)

Kinoshita,K: "Phase transition between hexagonal II (H_<II>) and liquid-cryscalline phase induced by interaction between solvents and segments of the membvane surface of dioleoylphsopha-tidylcholine." Biochim.Biophys.Acta. 1330. 199-206 (1997)

Kinoshita,K：“由溶剂和二油酰磷脂酰胆碱膜片表面部分之间的相互作用引起的六方 II (H_<II>) 和液晶相之间的相变。”

Kinoshita,K.: "Phase transition between hexagonal II (HII) and liquid-crystalline phase induced by interaction between solvents and segments of the membrane surface on dioleolyl phosphatidy lethano lamin" Biochimica et Biophysica Acta.1330. 199-206 (1997)

Kinoshita,K.：“由溶剂和二油酰磷脂酰乙醇核纤层蛋白上的膜表面片段之间的相互作用诱导的六方II (HII) 和液晶相之间的相变”Biochimica et Biophysicala Acta.1330。

Montal, O-M: "Formation of ion channels in lipid bilayers by a peptide with the predicted transmembrane sequence of botulinum neurotoxin A." Protein Science. vol.4. 1490-1497 (1995)

Montal, O-M：“通过具有预测的肉毒杆菌神经毒素 A 跨膜序列的肽在脂质双层中形成离子通道。”