Molecular dissection of organization and dynamics of lipid domains in biomembranes

生物膜中脂质结构域的组织和动力学的分子解剖

• 批准号: 17390025
• 负责人: KOBAYASHI Toshihide
• 金额: $ 9.6万
• 依托单位: RIKEN
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390025/
• 关键词: Biomembrane; Lipid bilayer; Lipid domain; Lipid probe; Lipid raft; Endocytosis; Endosome; Cholesterol; 後期エンドソーム; ビス(モノアシルグリセロ)リン酸; リゾビスボスファチジン酸; ガングリオシド; スフィンゴリピドーシス; 膜結合毒素; 膜ドメイン; スフィンゴミエリン; GM1; シグナル伝達

(1) Characterization of lipid probesLysenin is a pore-forming toxin that specifically binds sphingomyelin (SM). The binding of the toxin to the membrane is accompanied by the oligomerization of the protein, leading to pore formation. Although a previous study showed that SM/cholesterol liposomes were 10,000 times more effective than SM liposomes in inhibiting lysenin-induced hemolysis, the role of cholesterol is not precisely clarified. Our results indicate that both cholesterol and the SM/lysenin ratio control the amount of lysenin monomer via altering the state of protein oligomerization, thus affecting hemolysis.Duramycin is a 19 amino acid tetracyclic lantibiotic closely related to cinnamycin, which is known to bind phosphatidyl-ethanolamine (PE). The lipid specificity of duramycin was not established. Our study indicates that both duramycin and cinnamycin exclusively bind to ethanolamine phospholipids in curvature-dependent manner.(2) Regulation of endocytosis by cholesterolCellular cholesterol increases when cells reach confluency in Chinese hamster ovary (CHO) cells. We showed that the endocytic pathways of several fluorescent lipids were altered in cell confluency-dependent manner. The crucial role of cellular cholesterol in cell-confluency dependent endocytosis was suggested. Our results also suggest that cholesterol controls endocytic routes of a subset of membrane lipids through rab11.(3) Regulation of cellular cholesterol homeostasis by endosome lipid domainsUsing D-PDMP and ganglioside GM1, it is shown that the structure alteration of bis (monoacylglycero) phosphate (BMP)-rich late endosome lipid domain plays a crucial role in cellular cholesterol accumulation. We also employed anti-BMP monoclonal antibody to examine the role of BMP domain in cholesterol homeostasis in macrophages.

（1）脂质探针溶酶果的表征是一种特异性结合鞘磷脂（SM）的孔形成毒素。毒素与膜的结合伴随着蛋白质的寡聚，从而导致孔形成。尽管先前的研究表明，SM/胆固醇脂质体的有效性比SM脂质体在抑制溶烯蛋白诱导的溶血方面高10,000倍，但胆固醇的作用尚未精确阐明。我们的结果表明，胆固醇和SM/溶酶比通过改变蛋白质寡聚的状态来控制溶烯蛋白单体的量，从而影响溶血性。-二霉素是一种19氨基酸四乙酸四环lantaclic lantaibiotic，与肉桂蛋白密切相关，已知结合磷酸 - 乙醇氨基甲酰胺（PE）。未建立杜拉米霉素的脂质特异性。我们的研究表明，杜拉米霉素和肉桂素都以曲率依赖性的方式专门与乙醇胺磷脂结合。（2）当中国仓鼠卵巢（CHO）细胞中胆固醇胆固醇升高时，胆固醇细胞胆固醇通过胆固醇细胞胆固醇的增加而调节内吞作用。我们表明，几种荧光脂质的内吞途径在细胞汇合依赖性方式中发生了改变。提出了细胞胆固醇在细胞依赖性内吞作用中的关键作用。 Our results also suggest that cholesterol controls endocytic routes of a subset of membrane lipids through rab11.(3) Regulation of cellular cholesterol homeostasis by endosome lipid domainsUsing D-PDMP and ganglioside GM1, it is shown that the structure alteration of bis (monoacylglycero) phosphate (BMP)-rich late endosome lipid domain plays a在细胞胆固醇积累中的关键作用。我们还采用抗BMP单克隆抗体来检查BMP结构域在胆固醇稳态中的作用。

项目成果

pH-dependent formation of membranous cytoplasmic body-like structure of ganglioside GM1/bis(monoacylglycero)phosphate mixed membranes

• DOI: 10.1529/biophysj.106.098657
• 发表时间: 2007-01-01
• 期刊: BIOPHYSICAL JOURNAL
• 影响因子: 3.4
• 作者: Hayakawa, Tomohiro;Makino, Asami;Kobayashi, Toshihide
• 通讯作者: Kobayashi, Toshihide

Corrective effect of yeast recombinant human α-galactosidase having N-linked sugar chains suitable for lysosomal delivery on Fabry mice

适合溶酶体递送的具有N-连接糖链的酵母重组人α-半乳糖苷酶对法布里小鼠的纠正作用

• 发表时间: 2006
• 期刊: J Hum Genet (in press)
• 作者: Hayakawa T;Delton-Vandenbroucke I;Hayakawa T;Ishitsuka R;Matsunaga S;Hullin-Matsuda F;Sakuraba H;Hayakawa T;Singh RD;Makino A;Sakuraba H
• 通讯作者: Sakuraba H

D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol alters cellular cholesterol homeostasis by modulating the endosome lipid domains

• DOI: 10.1021/bi052104y
• 发表时间: 2006-04-11
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Makino, A;Ishii, K;Kobayashi, T
• 通讯作者: Kobayashi, T

Spatial and functional heterogeneity of sphinogolipid-rich membrane domains

富含鞘脂的膜域的空间和功能异质性

• 发表时间: 2005
• 期刊: J Biol Chem 280
• 作者: Hayakawa T;Delton-Vandenbroucke I;Hayakawa T;Ishitsuka R;Matsunaga S;Hullin-Matsuda F;Sakuraba H;Hayakawa T;Singh RD;Makino A;Sakuraba H;Kiyokawa E;Miyaji M;Kiyokawa E
• 通讯作者: Kiyokawa E

Spatial and functional heterogeneity of sphingolipid-rich membrane domains

• DOI: 10.1074/jbc.m502244200
• 发表时间: 2005-06-24
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Kiyokawa, E;Baba, T;Kobayashi, T
• 通讯作者: Kobayashi, T