Optically Modulated Fluorescent Proteins

光调制荧光蛋白

• 批准号: 7904789
• 负责人: ROBERT M DICKSON
• 金额: $ 17.93万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7904789
• 关键词: Autoimmune Diseases; Back; Biochemical Process; Blinking; Cells; Characteristics; Coupling; Defect; Disease; Electronics; Fluorescence; Frequencies; Image; Isomerism; Lasers; Life; Literature; Malignant Neoplasms; Molecular; Mutagenesis; Mutate; Nobel Prize; Noise; Optics; Output; Pathway interactions; Proteins; Protons; Public Health; Rest; Set protein; Signal Transduction; Site-Directed Mutagenesis; Techniques; Time; Tyrosine; absorption; base; computerized data processing; deprotonation; design; fluorophore; improved; irradiation; public health relevance; quantum; research study; residence; single molecule; stereochemistry

DESCRIPTION (provided by applicant): Advances in molecularly specific fluorescent imaging have revolutionized the tracking of biochemical processes in cells. A continuing problem, however, has been the relatively poor signal-to-noise ratio in the presence of background fluorescence from endogenous or exogenous fluorophores. We propose to employ a signal processing technique called optical modulation to improve that ratio. We will design and synthesize proteins based upon a blue fluorescent protein which will undergo fast conversion to a green-absorbing form. This form can be converted back to the fluorescent form by long- wavelength irradiation, producing a tracking signal which can be used for optical modulation and lock-in amplification. By mutating known blue fluorescent proteins or photoswitchable proteins toward the right absorption/isomerizations characteristics, we will develop optically modulatable fluorescent proteins capable of true intracellular single molecule imaging. PUBLIC HEALTH RELEVANCE: Public health statement Optically Modulated Fluorescent Proteins Fluorescent proteins have revolutionized the tracking of biochemical processes in cells, as recognized by the 2008 Nobel Prize in the field and by the presence of over 100,000 references to the use of fluorescent proteins in the scientific literature. Using new proteins derived from known fluorescent proteins, we propose to develop a new set of proteins, coupling them to a signal processing technique called optical modulation, to greatly improve sensitivity. The ability to follow the dynamics of cellular pathways at the near molecular level will allow significant progress to be made in disease states that are dependent upon defects in cell transport, including cancer and autoimmune diseases.

描述（由申请人提供）：分子特异性荧光成像的进步彻底改变了细胞中生化过程的追踪。然而，一个持续存在的问题是在内源或外源荧光团的背景荧光存在下信噪比相对较差。我们建议采用一种称为光调制的信号处理技术来提高该比率。我们将设计和合成基于蓝色荧光蛋白的蛋白质，该蛋白质将快速转化为吸收绿色的形式。通过长波长照射，这种形式可以转换回荧光形式，产生可用于光学调制和锁定放大的跟踪信号。通过将已知的蓝色荧光蛋白或光开关蛋白突变为正确的吸收/异构化特征，我们将开发能够真正进行细胞内单分子成像的光学可调节荧光蛋白。公共健康相关性：公共健康声明 光调制荧光蛋白 荧光蛋白彻底改变了细胞生化过程的追踪，这一点得到了 2008 年该领域诺贝尔奖的认可，并且有超过 100,000 篇关于荧光蛋白在科学领域中使用的参考文献所认可。文学。使用源自已知荧光蛋白的新蛋白质，我们建议开发一组新的蛋白质，将它们与称为光调制的信号处理技术耦合，以大大提高灵敏度。在近分子水平上跟踪细胞通路动态的能力将使依赖于细胞运输缺陷的疾病状态（包括癌症和自身免疫性疾病）取得重大进展。

项目成果

Green fluorescent protein with anionic tryptophan-based chromophore and long fluorescence lifetime.

绿色荧光蛋白，具有阴离子色氨酸发色团，荧光寿命长。

• DOI: 10.1016/j.bpj.2015.06.018
• 发表时间: 2015-07-21
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: K. Sarkisyan;A. S. Goryashchenko;Peter V. Lidsky;D. Gorbachev;N. Bozhanova;A. Gorokhovatsky;Alina Pereverzeva;Alina P. Ryumina;V. Zherdeva;A. Savitsky;K. M. Solntsev;A. Bommarius;G. Sharonov;Jake R. Lindquist;M. Drobizhev;T. Hughes;A. Rebane;K. Lukyanov;A. Mishin
• 通讯作者: A. Mishin

Fluorescence imaging using synthetic GFP chromophores.

使用合成 GFP 发色团进行荧光成像。

• DOI: 10.1016/j.cbpa.2015.06.002
• 发表时间: 2015-08-01
• 期刊: Current opinion in chemical biology
• 影响因子: 7.8
• 作者: C. Walker;K. Lukyanov;I. Yampolsky;A. Mishin;A. Bommarius;Anna M. Duraj;Bahareh Azizi;L. Tolbe
• 通讯作者: L. Tolbe

Hidden photoinduced reactivity of the blue fluorescent protein mKalama1.

蓝色荧光蛋白 mKalama1 的隐藏光诱导反应性。

• 发表时间: 2015-05-21
• 作者: Vegh, Russell B;Bloch, Dmitry A;Bommarius, Andreas S;Verkhovsky, Michael;Pletnev, Sergei;Iwaï, Hideo;Bochenkova, Anastasia V;Solntsev, Kyril M
• 通讯作者: Solntsev, Kyril M

Chromophore photoreduction in red fluorescent proteins is responsible for bleaching and phototoxicity.

红色荧光蛋白中发色团的光还原作用导致漂白和光毒性。

• 发表时间: 2014-05-01
• 作者: Vegh, Russell B;Bravaya, Ksenia B;Bloch, Dmitry A;Bommarius, Andreas S;Tolbert, Laren M;Verkhovsky, Michael;Krylov, Anna I;Solntsev, Kyril M
• 通讯作者: Solntsev, Kyril M

Optically modulatable blue fluorescent proteins.

可光调制的蓝色荧光蛋白。

• DOI: 10.1021/ja405459b
• 发表时间: 2013-11-06
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Jablonski AE;Vegh RB;Hsiang JC;Bommarius B;Chen YC;Solntsev KM;Bommarius AS;Tolbert LM;Dickson RM
• 通讯作者: Dickson RM