Molecular Bases on Bioluminescence and Luminous Behavior Professor

生物发光和发光行为的分子基础教授

• 批准号: 09041100
• 负责人: NIWA Haruki
• 金额: $ 7.04万
• 依托单位: The University of Electro-communications
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09041100/
• 关键词: New Zealand; luminous organism; Latianeritoides; luciferin; luciferase; m-RNA; c-DNA; cloning; Lafia; C-DNA; Arachnocampa; Latia; c-DNA; Octochaetus

Molecular base on bioluminescence of fresh water limpet-like snails Latia neritoides in New Zealand was investigated. Rapid and convenient method for purification of the Latia luciferase was established. (1) The Latia luciferase was found to be an N-linked glycoprotein, and to exhibit bioluminescence activity without any cofactor. The size exclusion chromatography analysis coupled with SDS-PAGE analysis and MALDI-TOF mass spectrometry indicated that the bioluminescent active Latia luciferase is a homohexamer, whose molecular weight is ca. 180,000 and the molecular weight of the non-bioluminescent subunit protein is ca. 31,000. (2) Although the natural Latia luciferin has an enol formate group, the formyl one was found to be not essential functionality, but enol acetate has also bioluminescence activity, indicating that the bioluminescence system of Latia is completely different from bacteria's ones believed previously and that the Latia luciferase has also an esterase activity. The formation of enolate anion in the Latia luciferase may be essential initial event in the bioluminescence reaction. (3) The Latia m-RNA was extracted from live specimens, from which Latia c-DNA library was constructed. Now cloning of the luciferase c-DNA is in progress.

研究了新西兰淡水帽贝类蜗牛 Latia neritoides 生物发光的分子基础。建立了快速、便捷的Latia荧光素酶纯化方法。 (1) Latia荧光素酶被发现是一种N-连接糖蛋白，并且在没有任何辅因子的情况下表现出生物发光活性。尺寸排阻色谱分析结合SDS-PAGE分析和MALDI-TOF质谱表明生物发光活性Latia荧光素酶是同源六聚体，其分子量约为1。 180,000，非生物发光亚基蛋白的分子量约为180,000。 31,000。 (2)虽然天然的Latia荧光素具有烯醇甲酸酯基，但发现甲酰基不是必需的功能，但烯醇乙酸酯也具有生物发光活性，这表明Latia的生物发光系统与之前认为的细菌的生物发光系统完全不同，并且Latia荧光素酶也具有酯酶活性。 Latia 荧光素酶中烯醇阴离子的形成可能是生物发光反应中必不可少的初始事件。 (3)从活体标本中提取Latia mRNA，构建Latia c-DNA文库。目前，荧光素酶 c-DNA 的克隆正在进行中。

项目成果

Satoshi Kojima, Hiroko Ohkawa, Takashi Hirano, Haruki Niwa, Shojiro Maki, Mamoru Ohashi, Satoshi Inouye, and Frederick I.Tsuji.: "Fluorescent Properties of Model Chromophores of Tyrosine-66 Substituted Mutants of Aequorea Green Fluorescent Protein (GFP)."

Satoshi Kojima、Hiroko Ohkawa、Takashi Hirano、Haruki Niwa、Shojiro Maki、Mamoru Ohashi、Satoshi Inouye 和 Frederick I.Tsuji.：“水母绿色荧光蛋白 (GFP) 酪氨酸 66 取代突变体模型发色团的荧光特性。”

Nobuyoshi Ohba: "Mating Behavior of the Firefly, Pyrocoelia matsumurai (Coleoptera ; Lampiridae)."Sci.Rept.Yokosuka City Mus.. 45. 45-50 (1997)

Nobuyoshi Ohba：“萤火虫的交配行为，Pyrocoelia msumurai（鞘翅目；萤科）。”Sci.Rept.Yokosuka City Mus.. 45. 45-50 (1997)

Toshikazu Hiraki, Masayasu Kubota, Takashi Hirano, Haruki Niwa, and Mamoru Ohashi: "FAB-CID Mass Spectrometry of Imizdazplone Derivatives Related to the Chromophore of Green Fluorescent Potein in Jellyfish Bioluminescence."J.Mass Spectrom.Jpn.. 46, No. 1.

Toshikazu Hiraki、Masayasu Kubota、Takashi Hirano、Haruki Niwa 和​​ Mamoru Ohashi：“与水母生物发光中绿色荧光蛋白发色团相关的咪唑啉衍生物的 FAB-CID 质谱法。”J.Mass Spectrom.Jpn.. 46，第 46 期。

Satoshi Kojima, Shojiro Maki, Takashi Hirano, Yoshihilo Oumiya, and Haruki Niwa: "Bioluminescence Activity of Latia Luciferin Analogos."Tetrohedron Lett.. 41, No. 22. 4409-4413 (2000)

Satoshi Kojima、Shojiro Maki、Takashi Hirano、Yoshihilo Oumiya 和 Haruki Niwa：“Latia Luciferin 类似物的生物发光活性。”Tettrohedron Lett.. 41，第 22 期。4409-4413 (2000)

J.J.Zheng et al.: "Synthesis and Chemi-and Bioluminesence Properties of a Photolabile Coelenterazine Analogue Bearing an Azido Group"Bull. Chem. Soc. Jpn.. 73. 465-469 (2000)

J.J.Zheng 等人：“带有叠氮基团的光不稳定腔肠素类似物的合成及其化学和生物发光特性”公牛。