The elucidation of the low-temperature stress mechanism using reconstituted vesicles with tonoplast H^+-ATPase

使用液泡膜 H^ -ATP 酶重建囊泡阐明低温应激机制

• 批准号: 11440237
• 负责人: KASAMO Kunihiro
• 金额: $ 9.09万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11440237/
• 关键词: Vacuoles; Proton pump; proteoliposomes; membrane fluidity; Proton translocating; low temperature stress; glycolipid; 膜流動性

Proton pumping across the tonoplast was markedly suppressed by chilling. In order to clarify the mechanism of chilling-induced depression of proton-pumping across tonoplast. we reconstituted chimera proteoliposomes with H^+-ATPase solubilized from the tonoplast of chilling insensitive rice (var. Nipponbare) without chilling and individual or combined lipids extracted from the tonoplast of chilling-sensitive rice (var. Boro) cells with or without chilling at 5 ℃. We further examined the relation between membrane fluidity and chilling.As the result, the chilling-induced decrease in proton pumping across the tonoplast is due to the decrease in the fluidity of the hydrophilic surface region of the lipid bilayer of the tonoplast, which is caused by glycolipids, such as glucocerebroside and acylsteryl glucoside.

质子泵送通过张力质体明显抑制冷却。为了阐明低温诱导的质子抽运抑制机制。用冷冻不敏感水稻（var. Nipponbare）的张力质体和冷冻不敏感水稻（var. Boro）的张力质体中提取的单个或组合脂质，在5℃条件下重组H^+- atp酶溶解的嵌合体蛋白脂质体。我们进一步研究了膜流动性与冷却的关系。因此，冷却引起的质子泵送穿过张力质体的减少是由于张力质体脂质双分子层亲水性表面区域流动性的减少，这是由糖脂类引起的，如糖脑苷和酰基乙酰葡萄糖苷。

项目成果

Kasamo, K.: "Mechanism of the chilling-induced decrease in proton pumping across the tonoplast of rice cells"Plant Cell Physiol.. 41. 840-849 (2000)

Kasamo, K.：“寒冷诱导的水稻细胞液泡质子泵送减少的机制”Plant Cell Physiol.. 41. 840-849 (2000)

Yamaguchi, M: "Modulation in the activity of purified tonoplast H^+-ATPase by tonoplast glycolipids prepared from cultured rice cells"Plant Cell Physiol.. 42. 516-523 (2001)

Yamaguchi，M：“通过从培养的水稻细胞制备的液泡膜糖脂调节纯化的液泡膜H + -ATP酶的活性”Plant Cell Physiol.. 42. 516-523 (2001)

Yamaguchi, M: "Modulation of proton-pumping across proteoliposome membrane reconstituted with tonoplast H^+-ATPase from cultured rice cells by acyl steryl glucoside"Plant Cell Physiol.. 43(in press). (2002)

Yamaguchi，M：“通过酰基甾基葡萄糖苷对用来自培养的水稻细胞的液泡膜H 2 -ATP酶重建的蛋白脂质体膜的质子泵送进行调节”植物细胞生理学.. 43（出版中）。

Yamaguchi,M.: "Modulation of the activity of purified tonoplast H^+-ATPase by tonoplast glycolopids prepared from cultured rice (Oryza sativa L.var Boro) cells."Plant & Cell physiol.. 42(印刷中). (2001)

Yamaguchi, M.：“通过从培养的水稻（Oryza sativa L.var Boro）细胞制备的液泡膜糖脂调节纯化的液泡膜H^+-ATP酶的活性。”植物与细胞生理学.. 42（出版中）。 ）