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Development of Fluorescent ON/OFF Mode Biosensor Based on GFP Chromophore

基于GFP发色团的荧光ON/OFF模式生物传感器的研制

基本信息

批准号：
12680588
负责人：
NIWA Haruki
金额：
$ 2.37万
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 Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

1) In connection with the development of new fluorescent biosensor based on GFP fluorescent chromophore several lead compounds with a 4-arylmethylidene-5-imidazolone structure were prepared and their fluorescence properties were investigated. Starting with 2-naphthalenecarboxaldehyde, 6-methoxy-2-naphthalenecarboxaldehyde, 4-(dimethylamino)benzaldehyde, and 1-pyrenecarboxaldehyde, we could prepare 1,2-dimethyl-4-(2-naphthylmethylidene)-5-imidazolone, 1,2-dimetnyl-4-(6-methoxy-2- naphthylmethylidene)-5-imidazolone, 1,2-dimethyl-4-[4-(dimethylamino)phenylmethylidene]-5-imidazolone, and 1,2-dimethyl-4-(1-pyrenylmethylidene)-5-imidazolone by utilizing Erlenmeyer ablactate method. We found that all compounds prepared exhibited very weak fluorescent properties in solution state, whereas they showed 2000〜3000 times strong fluorescence in ethanol glass matrices at 77 ° K. Fluorescence maximums were 520 nm for the 2-naphthyl derivative, 460nm for the 6-methoxy-2-naphthyl derivative, 520 nm for the 4-(dimethylamino)phenyl derivative, and 503 nm for the 1-pyrenyl derivative.2) We attempted to prepare the 4-aryliden-5-imidazolone derivative having metal-chelating functional groups and we could obtained 1-methyl-4-[(3-bromophenyl)methylidene]-2-(3-bromophenyl)-5-imidazolone, being able to introduce chelating groups. Reaction of the imidazolone derivatives with morpholine in the presence of a Pd(0) catalyst afforded the corresponding morpholino derivative. Introduction of a Ca-chelating functional group is under investigation.3) For studying fluorescence properties of the GFP chromophore, we investigated the relationship between fluorescence life times and solvent viscosities by using a GFP chromophore model imidazolone derivative. As results, the fluorescence lifetime of the GFP chromophore was prolonged with increasing the viscosity of the solvent used.

1)为了开发基于GFP荧光发色团的新型荧光生物传感器，制备了几种具有4-芳基亚甲基-5-咪唑啉酮结构的先导化合物，并研究了它们的荧光特性。以2-萘甲醛、6-甲氧基-2-萘甲醛、4-(二甲氨基)苯甲醛和1-芘甲醛为起始原料，我们可以制备1,2-二甲基-4-(2-萘基亚甲基)-5-咪唑啉酮， 1,2-二甲基-4-(6-甲氧基-2-萘基亚甲基)-5-咪唑酮、1,2-二甲基-4-[4-(二甲基氨基)苯基亚甲基]-5-咪唑酮和1,2-二甲基-4-(1-芘基亚甲基)-5-咪唑酮，通过使用Erlenmeyer 断奶法。我们发现制备的所有化合物在溶液状态下均表现出非常弱的荧光性能，而在77°K的乙醇玻璃基质中则显示出2000〜3000倍的强荧光。2-萘基衍生物的荧光最大值为520 nm，6-甲氧基-2-萘基衍生物为460 nm，4-(二甲氨基)苯基衍生物为520 nm，503 2)我们尝试制备具有金属螯合官能团的4-亚芳基-5-咪唑啉酮衍生物，得到了能够引入螯合基团的1-甲基-4-[(3-溴苯基)亚甲基]-2-(3-溴苯基)-5-咪唑啉酮。咪唑酮衍生物与吗啉在 Pd(0) 催化剂存在下反应得到相应的吗啉代衍生物。 Ca螯合官能团的引入正在研究中。3)为了研究GFP发色团的荧光特性，我们通过使用GFP发色团模型咪唑啉酮衍生物研究了荧光寿命和溶剂粘度之间的关系。结果，随着所用溶剂粘度的增加，GFP 发色团的荧光寿命延长。