Cell Biological Studies on Protein Body-Vacuole Transformation in Germinating Castor Bean Seedlings.

蓖麻子幼苗蛋白体-液泡转化的细胞生物学研究。

• 批准号: 62560079
• 负责人: NISHIMURA Mikio
• 金额: $ 1.34万
• 依托单位: Kobe 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: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560079/
• 关键词: Protein body; Vacuole; Membrane flow; Membrane fusion; Transformation of organelles; Castor bean; ヒマ; カボチャ; 11Sグロブリン; ヒマ種子; 貯蔵蛋白質; 特異抗体

Protein bodies are fused with each other and become finally a central vacuole during germination of fatty seed. We have analyzed the transfomation of protein bodies to vacuoles, especially focused on membrane fusion in membrane flow system.Methods for isolation of protein body membrane and vacuolar membrane from castor bean and pumpkin seed were established and the development of membrane proteins was studied. Four main membrane proteins (80 kD, 28 kD, 26 kD, 24 kd) were detected in protein body membrane from castor bean seed and amounts of these proteins were decreased during transformation of protein bodies to vacuoles. On the other hand, 5 main proteins (73 kD, 51 kD, 32 kD, 27 kD, 23 kD) were found in pumpkin protein body membrane. These membrane proteins were also decreased during the transformation whereas several new proteins (94 kD, 58 kD, 16 kD) appear in the membrane. Immunochemical analysis show that castor bean 80 kD and 26 kD were crossreacted with pumpkin 73 kD and 32 kD, suggesting that these proteins have similar structure and functions.We have prepared monospecific antibodies against these membrane proteins from pumpkin seed. The effect of these antibodies on fusion of protein body membrane are under experiments.

在种子萌发过程中，蛋白体相互融合，最终形成中央液泡。本文分析了蛋白体向液泡转化的过程，特别是膜流系统中的膜融合，建立了蓖麻籽和南瓜籽蛋白体膜和液泡膜的分离方法，并对膜蛋白的发育进行了研究。在蓖麻籽蛋白体膜上检测到4种主要的膜蛋白（80 kD、28 kD、26 kD、24 kD），在蛋白体向液泡转化过程中，这些蛋白的含量逐渐减少。在南瓜蛋白体膜上发现5种主要蛋白质（73 kD、51 kD、32 kD、27 kD、23 kD）。这些膜蛋白在转化过程中也有所减少，而在膜上出现了几个新的蛋白（94 kD，58 kD，16 kD）。免疫化学分析表明，蓖麻籽80 kD和26 kD蛋白与南瓜73 kD和32 kD蛋白具有交叉反应，表明这两种蛋白具有相似的结构和功能。这些抗体对蛋白体膜融合的影响正在实验中。

项目成果

Makoto Hayashi: "Nucleotide sequence of cloned cDNA coding for pumpkin 11S globulin subunit." Eur. J. Biochem.172. 627-632 (1988)

Makoto Hayashi：“编码南瓜 11S 球蛋白亚基的克隆 cDNA 的核苷酸序列。”

Makoto Hayashi: "Effect of monensin on intracellular transport and post-translational processing of pumpkin 11S globulin precursors in developing pumpkin cotyledons." FEBS Letters. 238. 197-200 (1988)

Makoto Hayashi：“莫能菌素对南瓜子叶发育过程中南瓜 11S 球蛋白前体的细胞内运输和翻译后加工的影响。”

林誠: FEBS Letters. 238. 197-200 (1988)

林诚：FEBS 快报238。197-200 (1988)

深澤朝子: Plant cell physiology. (1988)

深泽麻子：植物细胞生理学（1988）。

西村幹夫: "Mehtods in Enzymol.Vol.148"Preparation of protoplasts from plant tissues for organelle isolation" Academic Press, 27-34 (1987)

Mikio Nishimura：“Mehtods in Enzymol.Vol.148”从植物组织中制备原生质体用于细胞器分离”学术出版社，27-34（1987）