Biophysical studies of oncogenic Cdc42Hs constructs

致癌 Cdc42Hs 构建体的生物物理研究

• 批准号: 8103146
• 负责人: PAUL Damien ADAMS
• 金额: $ 14.63万
• 依托单位: UNIVERSITY OF ARKANSAS AT FAYETTEVILLE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-10 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8103146
• 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; GTP Binding; GTP-Binding Proteins; Goals; Guanine Nucleotide Dissociation Inhibitors; Guanine Nucleotides; 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 Dynamics; Protein Region; Proteins; 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; career development; cell transformation; cell type; chemotherapy; defined contribution; design; experience; genetic regulatory protein; guanine nucleotide binding protein; inhibitor/antagonist; innovation; member; metaplastic cell transformation; mutant; programs; protein protein interaction; protein structure; ras Proteins; rho; structural biology; success; therapeutic target; tool; tumorigenesis

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（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在多个信号转导途径中充当分子定时开关。该提案的重点是研究与其致癌潜力相关的 Cdc42Hs 形式的结构和动力学，并将为调节该活性的机制提供原子级信息。将要研究的 Cdc42Hs 构建体是 A) Cdc42Hs（无核苷酸）和 B) Cdc42Hs(F28L_?L8)。定义核苷酸耗尽形式的 Cdc42Hs 的结构和动力学将揭示 Cdc42Hs 的构象，该构象促进蛋白质效应子（例如刺激致癌活性的 Dbl）的结合。对 Cdc42Hs(F28L_?L8) 结构和动力学的研究应该揭示 Cdc42Hs 中相互作用的信息，这些相互作用阻断了单突变体 Cdc42Hs(F28L) 引起的致癌活性。 这项研究的直接目标是使用 NMR 和荧光作为结构技术，以更好地了解 Cdc42Hs 的变化及其与调节因子的相互作用影响导致癌症的各种重要信号转导途径的过程。这项研究的长期目标是更好地了解蛋白质结构和动力学如何与异常细胞功能相关，这将为未来研究开发合理的抗癌药物设计方案奠定基础。 Cdc42Hs 是 Ras 蛋白超家族的成员，该蛋白与结肠癌、乳腺癌和肺癌有关。这些研究将提供有关特定细胞信号传导过程的特定方面的信息，这些信息随后可能用于开发新的抗癌治疗方法，这些治疗方法具有特异性，并且比传统化疗方法的毒性可能更低。我拥有在生物分子研究中使用荧光光谱的背景，并使用核磁共振光谱来研究溶液中小肽的结构。该提案的指导阶段将为我提供获得尚未掌握的这些技术的几种使用经验的机会，这对于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，特别是信号转导领域拥有丰富的经验。这是这个项目的绝佳环境，也促进了我职业生涯的发展。本提案的研究生涯发展计划分两个阶段。第一阶段（指导）将允许研究 Cdc42Hs 的无核苷酸构建体。这一阶段的结构工具、数据分析程序和科学合作的经验将为成功过渡到第二阶段（独立）做好必要的准备，在此期间我将研究 Cdc42Hs(F28L_?L8) 的结构和动力学，从而进一步研究与癌症相关的生物分子的结构生物学。

项目成果

Prospective Development of Small Molecule Targets to Oncogenic Ras Proteins.

致癌 Ras 蛋白小分子靶点的前瞻性开发。

• DOI: 10.4236/ojbiphy.2013.34025
• 发表时间: 2013
• 期刊: Open journal of biophysics
• 作者: Chandrashekar,Reena;Adams,PaulD
• 通讯作者: Adams,PaulD

A switch I mutant of Cdc42 exhibits less conformational freedom.

• DOI: 10.1021/bi2004284
• 发表时间: 2011-07-19
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Chandrashekar, Reena;Salem, Omar;Krizova, Hana;McFeeters, Robert;Adams, Paul D.
• 通讯作者: Adams, Paul D.