Chemical Probes of Protein Prenylation

蛋白质异戊二烯化的化学探针

基本信息

批准号：
7228870
负责人：
RICHARD A GIBBS
金额：
$ 19.67万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-09-01 至 2008-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7228870
关键词：
Achievement Address Antineoplastic Agents Behavior Binding Biochemical Biological Biological Assay Boxing Cancer Cell Growth Cell model Cells Chemicals Clinical Trials Depth Development Dimethylallyltranstransferase Diphosphates Evaluation Event Exhibits Farnesol Future Glean Goals Grant Jurkat Cells Kinetics Laboratories Libraries Membrane Methods Modification Molecular Nature Numbers Oncogene Proteins Pathway interactions Pattern Peptide Library Peptides Post-Translational Protein Processing Principal Investigator Procedures Process Protein Isoforms Protein Isoprenylation Proteins Protocols documentation Relative (related person)Research Project Grants Role Screening procedure Signal Transduction Structure Synthetic Peptide Libraries Techniques Time Western Blotting Work analog base cancer therapy cell type cholesterol biosynthesis design inhibitor/antagonist insight interest intracellular protein transport isoprenoid mevalonate novel prenyl prenylation professor protein farnesyltransferase protein function protein geranylgeranyltransferase protein localization location ras Oncogene ras Proteins research study tool

项目摘要

DESCRIPTION (provided by applicant): Protein prenylation is a critically important post-translational modification process. It is required for the proper membrane association and activity of many signal transduction proteins, including the Ras oncogene products. There has been intense interest in the development of protein farnesyltransferase (FTase) inhibitors as anti-cancer agents, and promising results have been observed with these compounds in clinical trials. However, it is now clear that Ras is not the sole, or even the most important target of FTase inhibitors. The long-term goal of this research project is the development of chemical tools for the selective modulation of protein prenylation. These probes will allow for the determination of the relative roles of various prenylated proteins in cancer cell growth. This proposal will address the following specific hypothesis: The unique structure and mechanism of FTase allows for the discovery of selective inhibitors that will block the prenylation of certain proteins but not interfere with the prenylation of others. We will use currently existing and newly synthesized chemical tools, in combination with new biochemical and biological collaborative studies, to address the following specific aims: Aim 1) To characterize the interplay of peptide and isoprenoid selectivity in FTase and GGTase I. These studies will use existing and newly synthesized isoprenoid analogs, concise targeted synthetic peptide libraries, and various bioanalytical methods. Rational design will be employed along with screening methods to establish effective selectivity patterns. Aim 2) To elucidate the events covering selective prenylation by FTase at a molecular level, using existing kinetic techniques. Detailed steady state and pre-steady state kinetic experiments, in combination with various binding constant determinations, will be employed. These studies will provide novel insight into the mechanism employed by FTase, and insight into the design of new selective FPP analogs. Aim 3) To establish the ability of isoprenoid analogs to alter FTase substrate selectivity in cells, through the direct quantitation of prenyl isoforms of Ras and other prenylated proteins. The effects of farnesol analogs on flux through the mevalonate pathway and intracellularprenyl diphosphate pools also will be determined. The accomplishment of these aims will provide the tools and lay the groundwork for future studies to investigate the long-term hypothesis of this project: the selective introduction of modified prenyl groups into proteins will alter their subcellular localization and biological activity.

描述（由申请人提供）：蛋白质原始化是一个至关重要的翻译后修饰过程。适当的膜关联和许多信号转导蛋白的活性（包括RAS癌基产品）是必需的。人们对蛋白质Farnesylsylansferase（FTase）抑制剂作为抗癌剂的发展引起了极大的兴趣，在临床试验中，这些化合物在这些化合物中观察到了有希望的结果。但是，现在很明显，RAS不是FTase抑制剂的唯一或最重要的目标。该研究项目的长期目标是开发用于选择性调节蛋白质原始化的化学工具。这些探针将允许确定各种原蛋白在癌细胞生长中的相对作用。该提案将解决以下特定假设：FTase的独特结构和机制允许发现选择性抑制剂，这些抑制剂将阻止某些蛋白质的前化，但不会干扰其他蛋白质的前化。我们将使用当前现有和新合成的化学工具，再加上新的生物化学和生物协作研究，以解决以下具体目的：目标1）表征FTase和GGTase中肽和异磷酸化选择性的相互作用。这些研究将使用现有的和新的同步类似于类似的类似物，并构成了同步方法，并构成了同步方法，并将其使用。将采用理性设计以及筛选方法来建立有效的选择性模式。目的2）使用现有的动力学技术阐明涵盖通过分子水平的FTase选择性原苯化的事件。将采用详细的稳态和稳态的状态动力学实验，并结合各种结合常数确定。这些研究将提供有关FTase所采用的机制的新颖洞察力，并深入了解新的选择性FPP类似物的设计。目的3）通过直接定量Ras和其他原始化蛋白质的前定量，建立类似物类似物改变细胞中FTase底物选择性的能力。 Farnesol类似物对通过大甲酸盐途径和细胞内丙烯磷酸池的通量的影响也将得到确定。这些目标的完成将为未来的研究提供基础，以研究该项目的长期假设：选择性地引入修改后的蛋白质中的蛋白质将改变其亚细胞定位和生物学活性。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Synthesis of 7-substituted farnesyl diphosphate analogues.

7-取代法尼基二磷酸类似物的合成。

DOI：
10.1021/ol026176i
发表时间：
2002
期刊：
Organic letters
影响因子：
5.2
作者：
Rawat,DiwanS;Gibbs,RichardA
通讯作者：
Gibbs,RichardA

Computational and conformational evaluation of FTase alternative substrates: insight into a novel enzyme binding pocket.

FTase 替代底物的计算和构象评估：深入了解新型酶结合袋。

DOI：
10.1021/ci0496550
发表时间：
2005
期刊：
Journal of chemical information and modeling
影响因子：
5.6
作者：
Henriksen,BrianS;Zahn,ToddJ;Evanseck,JeffreyD;Firestine,StevenM;Gibbs,RichardA
通讯作者：
Gibbs,RichardA

Aromatic farnesyl diphosphate analogues: vinyl triflate-mediated synthesis and preliminary enzymatic evaluation.

芳香族法尼基二磷酸类似物：三氟甲磺酸乙烯酯介导的合成和初步酶促评价。

DOI：
10.1016/s0960-894x(02)00187-7
发表时间：
2002
期刊：
Bioorganic & medicinal chemistry letters
影响因子：
2.7
作者：
Zhou,Chunmei;Shao,Ying;Gibbs,RichardA
通讯作者：
Gibbs,RichardA

Structure, biological activity and membrane partitioning of analogs of the isoprenylated a-factor mating peptide of Saccharomyces cerevisiae.

酿酒酵母异戊二烯化 a 因子交配肽类似物的结构、生物活性和膜分配。

DOI：
10.1034/j.1399-3011.2000.00705.x
发表时间：
2000
期刊：
The journal of peptide research : official journal of the American Peptide Society
影响因子：
0
作者：
Xie,H;Becker,JM;Gibbs,RA;Naider,F
通讯作者：
Naider,F

Synthesis and biochemical evaluation of 3,7-disubstituted farnesyl diphosphate analogues.

3,7-二取代法呢基二磷酸类似物的合成和生化评价。