Biophysical studies of oncogenic Cdc42Hs constructs

致癌 Cdc42Hs 构建体的生物物理研究

基本信息

批准号：
7494160
负责人：
PAUL Damien ADAMS
金额：
$ 11.69万
依托单位：
UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-10 至 2012-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7494160
关键词：
Accounting Affect Affinity Amino Acids Antineoplastic Agents Area Behavior Binding Binding Proteins Biological Biological Process Breast Cell Proliferation Cell Signaling Process Cell physiology Chimera organism Collaborations Colon Data Data Analyses Defect Development Development Plans Dissociation Drug Design Event Exhibits Fluorescence Fluorescence Spectroscopy Forms Controls Fostering Foundations Future GTP Binding GTP-Binding Proteins Genus Cola Goals Guanine Nucleotide Dissociation Inhibitors Guanine Nucleotides Guanosine Diphosphate Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases HRAS gene Human Hydrolysis Investigation Lead Light Lung Malignant Neoplasms Mediating Mentors Methods Modeling Molecular Molecular Conformation Molecular Medicine Mutation NMR Spectroscopy Nature Neoplasm Metastasis Nucleotides Oncogenic Peptides Pharmaceutical Preparations Phase Phosphotransferases Preparation Procedures Process Protein Region Proteins Rate Relative (related person)Research Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Protein Solutions Specificity Structure Structure-Activity Relationship Techniques Testing Therapeutic Time Tumor Cell Invasion Universities Work base cancer cell cancer therapy career cell transformation cell type chemotherapy defined contribution design experience genetic regulatory protein guanine nucleotide binding protein inhibitor/antagonist innovation member metaplastic cell transformation mouse Gdi2 protein mutant programs protein protein interaction protein structure ras Proteins rho structural biology success therapeutic target tool tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): Cdc42Hs, a member of the Ras superfamily signal transduction proteins, binds guanine nucleotides and acts as a molecular timing switch in multiple signal transduction pathways. This proposal is centered around studying the structure and dynamics of Cdc42Hs forms associated with its oncogenic potential and will provide atomic level information into mechanisms that regulate this activity. The Cdc42Hs constructs that will be studied are, A) Cdc42Hs(nucleotide-free) and, B) Cdc42Hs(F28L_?L8). Defining the structure and dynamics of the nucleotide-depleted form of Cdc42Hs will shed light on the conformation(s) of Cdc42Hs that facilitate the binding of protein effectors, such as Dbl, that stimulate oncogenic activity. The study of the structure and dynamics of Cdc42Hs(F28L_?L8) should reveal information on interactions in Cdc42Hs that block oncogenic activity caused by the single mutant Cdc42Hs(F28L). The immediate goals of this research are to use NMR and fluorescence as structural techniques to gain a better understanding of the processes by which changes in Cdc42Hs and its interaction with regulators affect various important signal transduction pathways that lead to cancer. The long-term goals of this proposed research are to gain a better understanding of how protein structure and dynamics correlate with aberrant cell function, which will lay the foundation for future research into the development of a rational anti-cancer drug design program. Cdc42Hs is a member of the Ras superfamily of proteins, which have been implicated in cancers in the colon, breasts, and lungs. These studies will provide information on particular aspects of specific cell-signaling processes that may be subsequently used in the development of new anti-cancer treatments that are specific and may be less toxic than traditional chemotherapy approaches. I have a background in the use of fluorescence spectroscopy in the study of biological molecules and have used NMR spectroscopy to study structure of small peptides in solution. The mentored phase of this proposal will provide me with the opportunity to gain experience in several uses of these techniques that have not yet been mastered that will be crucial to my transition to independent investigation. The Department of Molecular Medicine at Cornell University has great experience in the area of applying physical methods to biological processes, and in particular, signal transduction. It is an excellent setting for this project, and also to foster the development of my career. The research career development plan of this proposal is two-phased. Phase I (Mentored) will allow for the study of the nucleotide-free construct of Cdc42Hs. The experience with the structural tools, data analysis procedures, and scientific collaboration in this phase will foster the necessary preparation for a successful transition into Phase II (Independent), during which time I will study the structure and dynamics of Cdc42Hs(F28L_?L8), leading to further research in the structural biology of biomolecules related to cancer.

描述（由申请人提供）：CDC42HS是RAS超家族信号转导蛋白的成员，结合鸟嘌呤核苷酸，并充当多个信号转导途径中的分子正时开关。该建议集中在研究与其致癌潜力相关的Cdc42hs形式的结构和动力学围绕，并将将原子水平的信息提供为调节该活动的机制。将研究的CDC42HS构建体为a）cdc42hs（无核苷酸）和b）cdc42hs（f28l_？l8）。定义cdc42hs核苷酸耗尽形式的结构和动力学将阐明Cdc42hs的构象，从而促进刺激致癌活性的蛋白质效应子（例如DBL）的结合。对CDC42HS（F28L_？L8）的结构和动力学的研究应揭示有关CDC42HS相互作用的信息，这些信息阻断了由单个突变体Cdc42hs（F28L）引起的致癌活性。这项研究的直接目标是将NMR和荧光用作结构技术，以更好地了解Cdc42hs变化的过程及其与调节器的相互作用会影响导致癌症的各种重要信号转导途径。这项拟议的研究的长期目标是更好地了解蛋白质结构和动力学与异常细胞功能的相关性，这将为未来的研究奠定基础，以研究合理的抗癌药物设计计划的发展。 CDC42HS是蛋白质RAS超家族的成员，蛋白质与结肠，乳房和肺的癌症有关。这些研究将提供有关特定细胞信号过程的特定方面的信息，这些过程随后可用于开发新的抗癌治疗方法，这些治疗是特定的，并且可能比传统的化学疗法方法更少毒性。我在生物分子的研究中使用荧光光谱的使用背景，并使用NMR光谱研究了溶液中小肽的结构。该提案的指导阶段将为我提供机会，以获得这些技术的多种用途经验，这些技术尚未掌握，这些技术对我向独立调查的过渡至关重要。康奈尔大学分子医学系在将物理方法应用于生物过程，尤其是信号转导方面具有丰富的经验。这是该项目的绝佳环境，也可以促进我的职业发展。该提案的研究职业发展计划是两步的。 I期（指导）将允许研究Cdc42hs的无核苷酸构建体。在此阶段的结构工具，数据分析程序和科学合作的经验将促进成功过渡到II期（独立）的必要准备，在此期间，我将研究CDC42HS（F28L_？l8）的结构和动态，从而导致对与癌症相关的生物分析结构生物学的进一步研究。