Artificial Model for Inter-cellular communications

细胞间通信的人工模型

• 批准号: 03650721
• 负责人: KINOSHITA Takatoshi
• 金额: $ 1.02万
• 依托单位: Nagoya Institute of Technology
• 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-03650721/
• 关键词: Intercellular Communications; Gap Junction Model; Amphiphilic Polypeptide; Monolayer Reaction Method; Lipid Bilayer Vesicles; Vesicular Adhesion; Intervesicular Transport; Vesicular Recognition; 両親媒性ポリペプチド; ジパルミトイルホスファチジルコリン; 2分子膜ベシクル

Intercellular communication is mediated in many tissues by a major intrinsic protein called gap junction. It has been suggested, from analysis of the sequence of the gap junction protein, that it traverses the lipid bilayer via a transmembrane amphiphilic helix, which is essential for the formation of an intercellular aqueous channel. In the present study, we applied "Monolayer reaction method" to the preparation of the polypeptide composed of two amphiphilic helices jointed with a hydrophilic spacer between them, and examined its activity for the formation of an inter-vesicular aqueous channel as a gap junction model.The starting material, MPrM, was obtained by polymerization of the N-carboxy anhydride of L-glutamic acid eta-methyl ester with 1,3-diamino 2-propanohol as an initiator in 1,2-dichloroethane. And then MPrM was saponified by using the monolayer reaction method to give the amphiphilic nature to MPrM, am-MPrM. It can be concluded, from CD spectra of am-MPrM in dipalmitoylphosphatidylcholine, gel filtration analysis and inter-vesicular transport of D_2O, that am-MPrM incorporates its two amphiphilic helices to the bilayer membrane of different vesicles, respectively, to induce the vesicular adhesion and then associates each other in the membranes to form an aqueous channel, through which the micro-injection of D_2O can be effectively made. The vesicular recognition ability of the polypeptide will be required to make more realistic model systems.

在许多组织中，细胞间通讯由一种称为间隙连接的主要内在蛋白介导。根据对差距连接蛋白序列的分析，有人认为它通过跨膜两亲性螺旋穿过脂质双层，这对于细胞间水通道的形成至关重要。本研究采用“单层反应法”制备了由两个两亲性螺旋和一个亲水性间隔物连接而成的多肽，并将其作为间隙连接模型，研究了其形成囊泡间水通道的活性。3-二氨基2-丙醇作为引发剂在1，2-二氯乙烷中。然后用单分子层反应法对MPrM进行皂化，得到两亲性的MPrM，am-MPrM。从am-MPrM在二棕榈酰磷脂酰胆碱中的圆二色光谱、凝胶过滤分析和D2 O的囊泡间转运可以看出，am-MPrM的两个两亲性螺旋分别与不同囊泡的双层膜结合，诱导囊泡粘附，并在膜内相互缔合形成水通道，通过该通道可以有效地进行D2 O的微注射。多肽的囊泡识别能力将需要制作更逼真的模型系统。

项目成果

T.Kinoshita, M.Higuchi, A.Takizawa, Y.Tsujita: "Inter-vesicular Communications by a Synthetic Amphiphilic Polypeptide" Chem. Lett.,. 431-434 (1993)

T.Kinoshita、M.Higuchi、A.Takizawa、Y.Tsujita：“合成两亲性多肽的囊泡间通讯”化学。

T. Kinoshita: "The Preparation and Functions of a Photoresponsive Artificial Lipid" Senryou to Yakuhin,. Vol.38, No.2. 28-38 (1993)

T. Kinoshita：“光响应人工脂质的制备和功能” Senryou to Yakuhin，。

M.Higuchi,T.Kinoshita,A,Takizawa,Y.Tsujita,K.Okouchi,N.Hattori: "Interaction between an Anionic Sequential Polypeptide and Anionic Bilayer Membrane" Polymer Journal. 23. 15-22 (1991)

M.Higuchi,T.Kinoshita,A,Takizawa,Y.Tsujita,K.Okouchi,N.Hattori：“阴离子序列多肽与阴离子双层膜之间的相互作用”聚合物杂志。

M.Higuchi, T.Kinoshita, A.Takizawa, Y.Tsujita, K.Okouchi, N.Hattori: "Interaction between an Anionic Sequential Polypeptide and Anionic Bilayer Membrane" Polymer Journal. 23. 15-22 (1991)

M.Higuchi、T.Kinoshita、A.Takizawa、Y.Tsujita、K.Okouchi、N.Hattori：“阴离子序列多肽与阴离子双层膜之间的相互作用”聚合物杂志。

T. Kinoshita, M. Higuchi, A. Takizawa, and Y. Tsujita: "Intervesicular Transport by a Synthetic Channel Peptide Consisting og Two Amphiphilic Helices" Rept. Prog. Polym. Phys., Japan. Vol.35. 555-558 (1992)

T. Kinoshita、M. Higuchi、A. Takizawa 和 Y. Tsujita：“由两个两亲螺旋组成的合成通道肽的囊间运输”Rept。