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Physiological role of the pyrenoids in the green alga Bryopsis maxima.

绿藻 Bryopsis maxima 中蛋白核的生理作用。

基本信息

批准号：
63540541
负责人：
OKADA Mitsumasa
金额：
$ 1.15万
依托单位：
TOHO UNIVERSITY, Faculty of Science
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63540541/
关键词：
Bryopsis maxima CO_2 Fixation Green Agae Immunocytology Photosynthesis Pyrenoid RuBisCO ルピスコスビスコ

项目摘要

Physiological role of pyrenoids in the marine alga, Bryopsis maxima was studies using several methods as follows.I. Immunocytochemical researches. (1) The localization of ribulose 1,5-bisphosphate carboxylase/oxygenase(RuBisCO) in chloroplasts was examined by immunological techniques. Three strains of monoclonal antibodies against the large subunit of RuBisCO were made for the study. Immunofluorescence and immunoenzymatic studies showed that the large subunit of RuBisCO was concentrated in pyrenoids and on the surface of starch grains surrounding the pyrenoids.(2) The concentration of RuBisCO in Pyrenoids decreased when the alga was kept in the dark or matured to eject the gametes.II Biochemical studies (1) Intact pyrenoid core matrix was isolated by diethyl ether treatment and sucrose density gradient centrifugation using 1.8 M phosphate buffer. The purified pyrenoids contained RuBisCO and more than 10 minor polypeptides. They also showed RuBisCO activity when solubilized(specific activity was 0.62 umol CO2 fixed per mg protein per min). (2) The pyrenoids showed NADH-nitrate reductase activity and contained nitrite. The specific activity was 0.024 umols NO2 formed per mg protein per min, which was 80 times greater than that in the crude chloroplasts extract.III. Molecular biological research The gene of the large subunit of RuBisCO was cloned and sequenced. The gene has a large (2.3 kbp) intron and the product was not subjected to the posttranslational processing.

本文采用以下几种方法研究了大真藓（Bryopsis maxima）类蛋白核的生理作用。免疫细胞化学研究。(1)用免疫学方法研究了核酮糖1，5-二磷酸羧化酶/加氧酶（RuBisCO）在叶绿体中的定位。制备了三株抗RuBisCO大亚基的单克隆抗体。免疫荧光和免疫酶的研究表明，大亚基RuBisCO集中在蛋白核和周围的蛋白核的淀粉粒表面。(2)RuBisCO在类蛋白核中的浓度下降时，在黑暗中保存或成熟，以排出gametes.II生化研究（1）完整的类蛋白核核心基质分离乙醚处理和蔗糖密度梯度离心使用1.8 M磷酸盐缓冲液。纯化的类蛋白核含有RuBisCO和10多个次要多肽。它们在溶解时也显示出RuBisCO活性（比活性为0.62 μ mol CO2固定/mg蛋白/min）。(2)类蛋白核具有NADH-硝酸还原酶活性，并含有亚硝酸盐。比活性为0.024 umols NO2/mg蛋白/min，是叶绿体粗提物的80倍。分子生物学研究克隆了RuBisCO大亚基基因并进行了序列测定。该基因有一个大的（2.3 kbp）内含子和产物不进行翻译后加工。