Clonable Nanoparticles

可克隆纳米颗粒

基本信息

批准号：
8502254
负责人：
Christopher Jeffries Ackerson
金额：
$ 20.67万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8502254
关键词：
Amino Acids Antibiotics Area Bacteria Biocompatible Materials Biological Biological Assay Biology Cell Extracts Cells Chemicals Chemistry Chimeric Proteins Codon Nucleotides Complement Complex Cryoelectron Microscopy Custom Development Diagnosis Disease Electron Microscope Electron Microscopy Electrons Environment Exploratory/Developmental Grant Ferritin Filament Freezing Funding Gene Expression Genotype Gold Green Fluorescent Proteins In Situ In Vitro Individual Ions Learning Length Libraries Light Microscope Link Literature Magnetic Resonance Imaging Magnetism Mediating Messenger RNA Metals Microscope Microscopy Microtubules Minor Modality Modification Molecular Nobel Prize Optics Organism Oxidation-Reduction Peptide Fragments Peptide Library Peptide Phage Display Library Peptides Phage Display Phenotype Photons Play Property Protein Fragment Proteins Public Health Reading Reporter Genes Resolution Ribosomes Role Sampling Shapes Site Solutions Staining method Stains Structure System Techniques Technology Testing Time Tissues Tomogram Transition Elements Tubulin Visual Work Yeasts analog base cell fixing directed evolution disilver oxide electron crystallography fluorescence microscope germanium oxide in vivo iron oxide light microscopy magnetite ferrosoferric oxide metal oxide nanoparticle novel particle phytochelatin protein aminoacid sequence research study small molecule success tomography

项目摘要

DESCRIPTION (provided by applicant): The objective of this exploratory (R21) proposal is to identify the most promising strategy for solving the contrast problem in electron microscopy. In all microscopes, whether illuminated by photons or electrons, biological tissues are essentially transparent. Green Fluorescent Protein (GFP) and related fluorescent proteins have complemented small molecule stains to essentially solve the contrast problem in optical microscopy. For electron microscopy, however, there are no 'clonable' contrast markers analogous to GFP. Herein we propose to explore several peptide or small protein based strategies for nucleating, catalyzing or mediating the formation of electron dense inorganic nanoparticles which may allow the localization of individual proteins in macromolecular and whole cell electron microscopy. The proposed work is significant for four areas of inquiry. First and most significantly, a robust clonable nanoparticle will revolutionize electron microscopy, making it significant for all forms of biological EM. Second, we will study FtsZ, the prokaryotic tubulin analog as an initial target. Information we learn about the intrcellular structure and distribution of FtsZ is significant for the development of antibiotics that target FtsZ. Third, we will, in finding novel peptides or small proteins that interact with metal ions or coordination complexes learn more about the important and sometimes poorly understood interactions of metals and biomolecules. Fourth just as GFP can act as a 'reporter gene' for reading optically for gene expression in tissues and organisms, a genetically encodable magnetic nanoparticle may also serve as a widely used 'reporter gene' for detecting gene expression by MRI or CT. The proposed work will proceed in three specific aims. All candidate 'clonable nanoparticle' peptides or proteins identified in these specific aims will be evaluated in vitro, in situ and in vvo in an FtsZ based structural electron microscopy assay. Aims one and two use the directed evolution techniques of phage display and ribosome display, respectively, to isolate peptide sequences capable of provoking the formation of magnetic iron oxide nanoparticles. Aim 2 is technically more challenging but is more likely to yield a universally useful clonable nanoparticle than Aim 1, which may yield clonable nanoparticles that function in situ and in vitro, but not in vivo. Aim 3 is to assay naturally occurring peptides, proteins and relevant protein fragments, as well as peptides isolated by others, for comparison to the peptides we isolate in aims 1 and 2. The proposal identifies why existing 'known' peptides and proteins fail to function as EM contrast markers, and suggests modifications that we may make to these known peptides and proteins to adapt them as EM contrast markers. The public health significance of the development of the proposed enabling technologies will derive from both more comprehensive structural information on normal and diseased cells, and also from greater understanding of FtsZ biology, which may enable development of new antibiotics.

描述（由申请人提供）：此探索性（R21）建议的目的是确定解决电子显微镜中对比度问题的最有希望的策略。在所有显微镜中，无论是由光子还是电子照明，生物组织本质上都是透明的。绿色荧光蛋白（GFP）和相关的荧光蛋白已经补充了小分子染色，可以在光学显微镜中解决对比度问题。然而，对于电子显微镜，没有类似于GFP的“可克隆”对比标记。本文中，我们建议探索几种肽或基于小蛋白质的策略，以成核，催化或介导电子致密无机纳米颗粒的形成，这可能允许单个蛋白质在大分子分子和全细胞电子显微镜中定位。拟议的工作对于四个调查领域很重要。首先，也是最重要的是，可靠的纳米颗粒将彻底改变电子显微镜，使其对所有形式的生物EM显着。其次，我们将研究FTSZ，核小管蛋白类似物作为初始靶标。我们了解有关FTSZ的细胞细胞结构和分布的信息对于靶向FTSZ的抗生素的发展很重要。第三，我们将找到与金属离子或协调复合物相互作用的新型肽或小蛋白质，了解更多有关金属和生物分子的相互作用的重要且有时了解不足的相互作用。就像GFP可以充当组织和生物体中基因表达的光学读取的“报告基因”一样，遗传编码的磁性纳米颗粒也可以用作广泛使用的“报告基因”，用于通过MRI或CT检测基因表达。拟议的工作将以三个特定的目标进行。在基于FTSZ的结构电子显微镜测定法中，将评估所有特定目标中确定的所有候选纳米颗粒肽或蛋白质。目标和第二分别使用噬菌体显示和核糖体显示的定向进化技术，以分离能够引起磁性铁氧化铁纳米颗粒形成的肽序列。 AIM 2从技术上讲是更具挑战性的，但更有可能产生普遍有用的可克隆纳米颗粒比AIM 1可能产生原位和体外功能的可克隆纳米颗粒，但不能产生体内。 Aim 3 is to assay naturally occurring peptides, proteins and relevant protein fragments, as well as peptides isolated by others, for comparison to the peptides we isolate in aims 1 and 2. The proposal identifies why existing 'known' peptides and proteins fail to function as EM contrast markers, and suggests modifications that we may make to these known peptides and proteins to adapt them as EM contrast markers. 所提出的促成技术发展的公共卫生意义将既来自正常细胞和患病细胞的更全面的结构信息，又要源于对FTSZ生物学的更多了解，这可能使新的抗生素发展。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（2）

Sensitive, selective analysis of selenium oxoanions using microchip electrophoresis with contact conductivity detection.

DOI：
10.1021/ac502013k
发表时间：
2014-08-19
期刊：
ANALYTICAL CHEMISTRY
影响因子：
7.4
作者：
Noblitt, Scott D.;Staicu, Lucian C.;Ackerson, Christopher J.;Henry, Charles S.
通讯作者：
Henry, Charles S.

Radicals Are Required for Thiol Etching of Gold Particles.

DOI：
10.1002/anie.201502934
发表时间：
2015-08-03
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
0
作者：
Dreier TA;Ackerson CJ
通讯作者：
Ackerson CJ

Radiofrequency heating pathways for gold nanoparticles.

DOI：
10.1039/c4nr00464g
发表时间：
2014-08-07
期刊：
Nanoscale
影响因子：
6.7
作者：
Collins CB;McCoy RS;Ackerson BJ;Collins GJ;Ackerson CJ
通讯作者：
Ackerson CJ

Conformation and dynamics of the ligand shell of a water-soluble Au102 nanoparticle.

DOI：
10.1038/ncomms10401
发表时间：
2016-01-21
期刊：
Nature communications
影响因子：
16.6
作者：
Salorinne K;Malola S;Wong OA;Rithner CD;Chen X;Ackerson CJ;Häkkinen H
通讯作者：
Häkkinen H

Polymorphism in magic-sized Au144(SR)60 clusters.

DOI：
10.1038/ncomms11859
发表时间：
2016-06-14
期刊：
Nature communications
影响因子：
16.6
作者：
Jensen KM;Juhas P;Tofanelli MA;Heinecke CL;Vaughan G;Ackerson CJ;Billinge SJ
通讯作者：
Billinge SJ