ダイズ貯蔵タンパク質グリシニンの液胞へのターゲティングシグナルの解析

大豆贮藏蛋白甘氨酸对液泡的靶向信号分析

• 批准号: 09760132
• 负责人: 竹中 康之
• 金额: $ 1.34万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-09760132/
• 关键词: Glycinin; Proglycinin; Crystallization; Structure; Targeting

本研究は、プログリシニン及び成熟型グリシニンの高次構造を比較することにより、グリシニンの液胞へのターゲティングシグナノレ構造を明らかにすることを目的としている。本年度は、以下の点について朋究を行った。1. プログリシニンの構造解析プログリシニンの構造を解くにあたって、多重重原子同型置換法を用いた。昨年度では、15から3Aまでの回折データを用いて行った精密化は、R値が22.7%であった。本年度では、さらにモデリングと精密化を繰り返すことにより、R値が20.0%、free-Rが25.9%となり、プログリシニンの構造解折を完成させた2. 成熟型グリシニンの構造解析成熟型グリシニンは、NES緩衝液(pH5.8)中で、沈澱剤としてMPDを使用し、20℃で6ケ月間保温したところ、X線回折実験に充分な大きさを持つ単結晶(0.3×0.3×0.3mm)を得た。本結晶の予備的X線結晶構造解析を行ったところ、本結晶は空間群R3、格子定数a=115,b=115,c=190Å α=β:90°,γ=120°であった。非対称単位中に2つのサブユニットを含んでいた。3. 今後の研究の展開成熟型グリシニンの結晶に関して2.5Å以上の回折強度を測定する。そこで、完成したプログリシニンの高次構造をサーチモデルとして分子置換去を試みる。分子置換方を適用できない場合は重原子同型置換法を用い、成熟型グリシニンの高次構造を解明する。そして、プログリシニン、成熟型グリシニンの高次構造を比較することにより、液胞へのターゲティングシグナルとして機能している領域構造を推定する。

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项目成果

T.Katsube.et al.: "N-Glycosylation does not affect assembly and targeting of proglycinin in yeast" Biochimica et Biophysica Acta. 1379. 107-117 (1998)

T.Katsube.等人：“N-糖基化不会影响酵母中甘氨酸原的组装和靶向”Biochimica et Biophysicala Acta。

T.Katsube.et al.: "N-Glycosylation dose not affect assembly and targeting of proglycinin in yeast" Biochimica et Biophsica Acta. 1379. 107-117 (1998)

T.Katsube. 等人：“N-糖基化不会影响酵母中甘氨酸原的组装和靶向”Biochimica et Biophsica Acta。