Modified Photoproteins as Labels and Molecular Switches in Bioanalysis

修饰光蛋白作为生物分析中的标签和分子开关

基本信息

批准号：
8729792
负责人：
Sylvia Daunert
金额：
$ 11.54万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-01-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8729792
关键词：
Aequorin Affect Amber Amino Acid Sequence Amino Acids Apoproteins Binding Binding Proteins Biological Biological Assay Biological Markers Bioluminescence Biomedical Engineering Biosensing Techniques Biotechnology Categories Cells Characteristics Codon Nucleotides Computer Analysis Detection Development DsRed Electronics Engineering Environment Family Funding Genetic Code Genetic Engineering Goals Hydrogen Bonding Image In Vitro Knowledge Label Lead Libraries Life Ligands Liquid substance Luciferases Luminescent Proteins Methods Microfluidics Molecular Molecular Biology Nanotechnology Optics Peptide Sequence Determination Performance Physiological Play Post-Translational Protein Processing Preparation Property Protein Binding Domain Proteins Protocols documentation Research Resolution Role Sampling Scaffolding Protein Scheme Scientist Series Signal Transduction Site Solutions Source Structure System Techniques Technology Time Transfer RNA Variant Work analog assay development base chromophore coelenterazine computer studies design directed evolution hybrid protein in vivo luminescence molecular recognition mutant nanoscale nanosensors obelin protein structure response scaffold tool

项目摘要

DESCRIPTION (provided by applicant): Continuous discoveries in bioengineering and more efficient molecular biology methods have allowed scientists to create new designer biomolecules with unique and distinct properties. In that regard, bio- nanotechnology and nanoscale analysis are becoming increasingly prevalent, raising the demand for techniques with high sensitivity that can detect biomolecules in biological samples. Bioluminescent photoproteins, such as aequorin, possess great potential to provide with a solution to this new challenge because they can be detected at low concentrations, and have different emission wavelengths depending on the protein variant used, a property that can be exploited in multiplex analysis. We now propose to prepare new aequorin variants to broaden the scope of their use in bioanalysis, thus allowing for detection of biomolecules that are not detectable by other technologies. Protein molecular switches with optical properties are another type of designer biomolecules that, in the presence of a target ligand, demonstrate an altered response manifested by an "on/off" signal. These molecules can be useful in a variety of applications, such as in the development of nanosensors for in vitro and in vivo detection. To that end, we plan to design and develop bioluminescent molecular switches that incorporate the recognition properties of binding proteins with the bioluminescence afforded by the aequorin variants. The hypotheses formulated for the proposed work are based on knowledge gained during our current funding period, and investigate the use of a series of computational and synthetic approaches along with genetic engineering strategies targeting the alteration of the electronic environment of the chromophore that should lead to new bioluminescent proteins and molecular switches with a wide range of spectral properties. These photoproteins will be employed in the development of assays for important biomolecules. Finally, we will investigate the use of the newly prepared bioluminescent molecular switches in the multiplex analysis of biomolecules and in the simultaneous analysis of molecules in single cells. We anticipate that the new photoproteins will provide with new enabling technologies for in vitro and in vivo biosensing, imaging, and multiplex analysis that have a number of advantages over existing methods.

描述（由申请人提供）：生物工程和更有效的分子生物学方法的不断发现使科学家能够创造出具有独特和独特特性的新设计生物分子。在这方面，生物纳米技术和纳米级分析变得越来越普遍，提高了对能够检测生物样品中生物分子的高灵敏度技术的需求。生物发光蛋白，例如水母发光蛋白，具有为这一新挑战提供解决方案的巨大潜力，因为它们可以在低浓度下被检测到，并且根据所使用的蛋白质变体具有不同的发射波长，这一特性可以在多重分析中利用。我们现在建议制备新的水母发光蛋白变体，以扩大其在生物分析中的使用范围，从而允许检测其他技术无法检测到的生物分子。具有光学特性的蛋白质分子开关是另一种类型的设计生物分子，在存在目标配体的情况下，表现出通过“开/关”信号表现出的改变的响应。这些分子可用于多种应用，例如开发用于体外和体内检测的纳米传感器。为此，我们计划设计和开发生物发光分子开关，将结合蛋白的识别特性与水母发光蛋白变体提供的生物发光结合起来。为拟议工作制定的假设基于我们当前资助期间获得的知识，并研究了一系列计算和合成方法的使用以及针对发色团电子环境改变的基因工程策略，这将导致新的结果具有广泛光谱特性的生物发光蛋白质和分子开关。这些发光蛋白将用于开发重要生物分子的检测方法。最后，我们将研究新制备的生物发光分子开关在生物分子多重分析和单细胞分子同步分析中的应用。我们预计新的发光蛋白将为体外和体内生物传感、成像和多重分析提供新的使能技术，这些技术比现有方法具有许多优势。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Highly Sensitive and Selective Direct Detection of Zika Virus Particles in Human Bodily Fluids for Accurate Early Diagnosis of Infection.

高灵敏度和选择性直接检测人体体液中的寨卡病毒颗粒，以实现感染的准确早期诊断。

DOI：
发表时间：
2019-04-30
期刊：
影响因子：
4.1
作者：
Pawley, Devon C;Ricciardi, Michael J;Dikici, Emre;Deo, Sapna K;Daunert, Sylvia
通讯作者：
Daunert, Sylvia

Beyond Antibodies as Binding Partners: The Role of Antibody Mimetics in Bioanalysis.

超越抗体作为结合伙伴：抗体模拟物在生物分析中的作用。

DOI：
10.1146/annurev-anchem-061516-045205
发表时间：
2017-06-12
期刊：
Annual review of analytical chemistry
影响因子：
0
作者：
Xiaowen Yu;Yu;E. Dikici;S. Deo;S. Daunert
通讯作者：
S. Daunert

Discrete event simulation model of sudden cardiac death predicts high impact of preventive interventions.

心源性猝死的离散事件模拟模型预测了预防性干预措施的巨大影响。

DOI：
发表时间：
2013
期刊：
影响因子：
4.6
作者：
Andreev, Victor P;Head, Trajen;Johnson, Neil;Deo, Sapna K;Daunert, Sylvia;Goldschmidt
通讯作者：
Goldschmidt

Directing and Potentiating Stem Cell-Mediated Angiogenesis and Tissue Repair by Cell Surface E-Selectin Coating.

通过细胞表面 E-选择素涂层指导和增强干细胞介导的血管生成和组织修复。

DOI：
发表时间：
2016
期刊：
影响因子：
3.7
作者：
Liu, Zhao;Daftarian, Pirouz;Kovalski, Letícia;Wang, Bo;Tian, Runxia;Castilla, Diego M;Dikici, Emre;Perez, Victor L;Deo, Sapna;Daunert, Sylvia;Velazquez, Omaida C
通讯作者：
Velazquez, Omaida C

Nanotechnology-Driven Therapeutic Interventions in Wound Healing: Potential Uses and Applications.

纳米技术驱动的伤口愈合治疗干预：潜在用途和应用。

DOI：
发表时间：
2017-03-22
期刊：
影响因子：
18.2
作者：
Hamdan, Suzana;Pastar, Irena;Drakulich, Stefan;Dikici, Emre;Tomic;Deo, Sapna;Daunert, Sylvia
通讯作者：
Daunert, Sylvia