Analysis of Protein-Ligand Binding on the Proteomic Scale

蛋白质组规模上的蛋白质-配体结合分析

基本信息

批准号：
9313895
负责人：
Michael C Fitzgerald
金额：
$ 33.25万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9313895
关键词：
4-Hydroxy-Tamoxifen Age Aging Alpha Cell Binding Binding Proteins Bioinformatics Biological Biological Process Biology Brain Breast Cancer Cell Breast Cancer Model Breast Cancer cell line Cell Culture Techniques Cell Line Chemicals Computers Data Data Analyses Diagnosis Disease Drug effect disorder Estrogen receptor positive Gene Expression Gene Expression Profiling Genome Heart Investigation Kidney Left MCF10A cells MCF7 cell MDA MB 231 MDA-MB-468 Manuals Mass Spectrum Analysis Measurement Methods Modeling Modification Molecular Molecular Profiling Mouse Protein Mus Pathway interactions Peptides Pharmaceutical Preparations Physiological Processes Protein Analysis Proteins Proteome Proteomics Research Series Statistical Methods Tamoxifen Techniques Therapeutic Agents Thermodynamics Tissue-Specific Gene Expression Tissues Work age related aged base biophysical techniques experimental study human disease improved malignant breast neoplasm mouse model protein biomarkers protein expression protein folding protein function protein profiling public health relevance therapeutic target tool

项目摘要

DESCRIPTION (provided by applicant): Genome- and proteome-based gene expression profiling studies have been widely used over the past decade to characterize biological states and drug action. While such differential gene expression profiling studies can help identify the cellular pathways and physiological processes associated with a biological state or with the biological activity of a therapeutic agent, they often fail to produce a detailed molecular level understanding of the biological state or drug mode-of-action because gene expression levels are not directly tied to protein function. This has limited the number of useful protein biomarkers and therapeutic targets discovered from such studies, and left gaps in our understanding of drug-action. Proposed here is the use of thermodynamic measurements of protein stability to profile the protein-interaction networks associated with different biological states and drug action. Such thermodynamic measurements of protein stability are expected to be more closely related to protein function than are protein expression levels, and thus produce more useful protein biomarkers and therapeutic targets of disease and generate a better understanding of drug action. The proposed work will use a chemical modification- and mass spectrometry-based method, termed SPROX, to make thermodynamic measurements of protein stability on the proteomic scale in several different age-, disease-, and drug-related biological states. The specific aims of this work are: (1) to streamline and improve the data analysis pipeline in proteome-wide SPROX experiments; (2) to characterize the thermodynamic stability of proteins derived from four different cell culture models of breast cancer using the SPROX technique and determine which proteins have altered stabilities in the different cell culture models; (3) to characterize the thermodynamic stability of proteins derived from breast cancer cells grown in the presence and in the absence of tamoxifen using the SPROX technique and determine which proteins have altered stabilities in the presence of tamoxifen; (4) to characterize the thermodynamic stability of mouse proteins derived from a mouse model of aging using the SPROX technique and determine which proteins have altered stabilities as a function of age; and (5) to use the proteins identified in (2)-(4) to characterize the altered protein interaction networks associated with the different age-, disease-, and drug-related biological states in this study.

描述（由申请人提供）：在过去的十年中，基于基因组和蛋白质组基因表达分析研究已被广泛使用，以表征生物态和药物作用。尽管这种差异基因表达分析研究可以帮助鉴定与生物学状态或治疗剂的生物学活性相关的细胞途径和生理过程，但它们通常无法产生对生物学状态或药物作用的详细分子水平的理解，因为基因表达水平与蛋白质的功能无关。这限制了有用的蛋白质生物标志物的数量以及从此类研究中发现的治疗靶标，并在我们对药物行动的理解中留下了差距。这里提出的是使用蛋白质稳定性的热力学测量值介绍与不同生物态和药物作用相关的蛋白质交互网络。与蛋白质表达水平相比，预期蛋白质稳定性的热力学测量与蛋白质功能更紧密相关，因此产生了更有用的蛋白质生物标志物和疾病的治疗靶标，并对药物作用有更好的了解。拟议的工作将使用称为SPROX的化学修饰和质谱法，以在几种不同的年龄，疾病和药物相关的生物学状态下对蛋白质组量表上蛋白质量的稳定性进行热力学测量。这项工作的具体目的是：（1）简化和改善全蛋白质组SPROX实验中的数据分析管道；（2）使用SPROX技术来表征蛋白质的热力学稳定性，这些蛋白质源自乳腺癌的四种不同细胞培养模型，并确定哪些蛋白质在不同的细胞培养模型中改变了稳定性；（3）表征源自在存在的乳腺癌细胞和使用SPROX技术的情况下生长的乳腺癌细胞的蛋白质的热力学稳定性，并确定哪些蛋白质在他莫昔芬存在下具有改变稳定性。（4）表征使用SPROX技术源自衰老的小鼠模型的小鼠蛋白的热力学稳定性，并确定哪些蛋白质随着年龄的函数改变了稳定性；（5）在本研究中使用（2） - （4）中鉴定的蛋白质来表征与不同年龄，疾病和药物相关的生物态相关的改变的蛋白质相互作用网络。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Are allergens more abundant and/or more stable than other proteins in pollens and dust?

DOI：
10.1111/all.14121
发表时间：
2019-12-10
期刊：
ALLERGY
影响因子：
12.4
作者：
Cabrera, Aurora;Randall, Thomas A.;Mueller, Geoffrey A.
通讯作者：
Mueller, Geoffrey A.

Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using SILAC-SPROX.

DOI：
10.1021/acs.jproteome.7b00283
发表时间：
2017-09-01
期刊：
Journal of proteome research
影响因子：
4.4
作者：
Liu F;Meng H;Fitzgerald MC
通讯作者：
Fitzgerald MC

Large-Scale Analysis of Breast Cancer-Related Conformational Changes in Proteins Using Limited Proteolysis.

DOI：
10.1021/acs.jproteome.6b00755
发表时间：
2016-12-02
期刊：
Journal of proteome research
影响因子：
4.4
作者：
Liu F;Fitzgerald MC
通讯作者：
Fitzgerald MC

Discovery of Tamoxifen and N-Desmethyl Tamoxifen Protein Targets in MCF-7 Cells Using Large-Scale Protein Folding and Stability Measurements.

DOI：
10.1021/acs.jproteome.7b00442
发表时间：
2017-11-03
期刊：
Journal of proteome research
影响因子：
4.4
作者：
Ogburn RN;Jin L;Meng H;Fitzgerald MC
通讯作者：
Fitzgerald MC

Discovery of Manassantin A Protein Targets Using Large-Scale Protein Folding and Stability Measurements.

DOI：
10.1021/acs.jproteome.6b00237
发表时间：
2016-08-05
期刊：
Journal of proteome research
影响因子：
4.4
作者：
Geer Wallace MA;Kwon DY;Weitzel DH;Lee CT;Stephenson TN;Chi JT;Mook RA Jr;Dewhirst MW;Hong J;Fitzgerald MC
通讯作者：
Fitzgerald MC