Structure and function of a photoconvertible Water-soluble chlorophyll protein from Chenopodium album.

来自藜的光转换水溶性叶绿素蛋白的结构和功能。

• 批准号: 14540602
• 负责人: SATOH Hiroyuki
• 金额: $ 2.37万
• 依托单位: Toho University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14540602/
• 关键词: Chlorophyll; WSCP; water-soluble chlorophyll protein; Chenopodium album; シロイヌナズナ; transgenic plant; CP668; CP740; 立体構造; Arabidopsis; シグナル配列; プロモーター解析; GFP

Participation of water-soluble chlorphyll binding proteins during chlorophyll biosynthesis, transportation, and/or its degradation processes have been predicted. However, such studies have not been performed extensively. In this study, we purified the water-soluble chlorophyll protein(WSCP) from Chenopodium album and its cDNA was cloned.Sequence analysis of its cDNA revealed that the WSCP possess N-terminal signal sequence, mature-WSCP of 169 residues including 22 residues of C-terminal processed region. Homology analysis revealed that the WSCP is a member of DUF538 family that is a family wide spread in green plant.The WSCP was express in E.coli. The recombinant WSCP was fractionated into insoluble fraction. Denaturing of the protein with urea and following renaturation was performed. We found a acceptable condition for the recovery of its chlorophyll binding and photo-converting activities.The poly clonal antibody was raised aganst the E.coli-expressed insoluble WSCP. Tissue and subcellular localization of WSCP were analyzed. The WSCP located in water-soluble fraction and thylakoid membrane fraction of chloroplast indicating that existance of membrane-bound form.To investigate its physiological function, the transgenic Arabidopsis expressing WSCP were produced. Phenotype of the transgenic plants is partial sterile, slow glowth, slow entering into reproductive growth phase.

水溶性叶绿素结合蛋白参与了叶绿素的生物合成、运输和/或降解过程。然而，这类研究尚未广泛开展。本研究从Chenopodium相册中纯化了水溶性叶绿素蛋白（water-soluble叶绿素protein， WSCP），并克隆了其cDNA。序列分析表明，该WSCP具有n端信号序列，成熟WSCP有169个残基，其中c端加工区有22个残基。同源性分析表明，WSCP是广泛分布于绿色植物中的DUF538家族的一个成员。WSCP在大肠杆菌中表达。重组WSCP分离成不溶性组分。用尿素使蛋白质变性，然后再进行变性。我们找到了恢复其叶绿素结合和光转化活性的可接受条件。建立了针对大肠杆菌表达的不溶性WSCP的多克隆抗体。分析WSCP的组织和亚细胞定位。WSCP位于叶绿体的水溶性部分和类囊体膜部分，表明存在膜结合形式。为了研究其生理功能，制备了表达WSCP的转基因拟南芥。转基因植株的表型为部分不育，生长缓慢，进入生殖生长期缓慢。

项目成果

D.Horigome et al.: "Purification, crystallization and preliminary X-ray analysis of a water-soluble chlorophyll protein from Brassica oleracea"Acta Crystallographica section D. D59. 2283-2285 (2003)

D.Horigome 等人：“来自甘蓝的水溶性叶绿素蛋白的纯化、结晶和初步 X 射线分析”《结晶学报》D.D59 部分。

Quantification of Minimal Residual Disease in Patients with e1a2 BCR-ABL Positive Acute Lymphoblastic Leukemia Using a Real-Time RT-PCR Assay.

使用实时 RT-PCR 检测对 e1a2 BCR-ABL 阳性急性淋巴细胞白血病患者的微小残留疾病进行定量。

• 发表时间: 2002
• 期刊: Leukimia 16
• 作者: Hiromitsu Yokota;N.H.Tsuno;Y.Tanaka;T.Fukui;K.Kitamura;H.Hirai;K.Osumi;N.Itou;H.Satoh;M.Okada;K.Nakahara
• 通讯作者: K.Nakahara

遺伝子組換え植物の医薬品・食品分野への応用

转基因植物在医药、食品领域的应用

• 发表时间: 2005
• 期刊: Medical Technology 33(4)
• 作者: 佐藤 浩之

C.Reinbothe et al.: "A novel role of water-soluble chlorophyll proteins in the transitory storage of chorophyllide"Plant Physiology. 134. 1-11 (2004)

C.Reinbothe 等人：“水溶性叶绿素蛋白在叶绿素短暂储存中的新作用”植物生理学。

Purification, Crystallization and Preliminary X-ray Analysis of a Water-soluble Chlorophyll Protein from Brassica oleracea L.var.acephala (kale).

来自甘蓝（羽衣甘蓝）的水溶性叶绿素蛋白的纯化、结晶和初步 X 射线分析。

• 发表时间: 2003
• 期刊: Acta Crystallographica Section D 59
• 作者: Daisuke Horigome;Hiroyuki Satoh;Akira Uchida
• 通讯作者: Akira Uchida