权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Investigation of the design, structure and mechanism of Mena protein interaction inhibitors

Mena蛋白相互作用抑制剂的设计、结构和机制研究

基本信息

批准号：
10408668
负责人：
Jackson Halpin
金额：
$ 6.98万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10408668
关键词：
Actins Affinity Behavior Binding Biological Assay Biology Biophysics Blood Circulation Body part C-terminal Cancer Patient Cell Culture Techniques Cells Cessation of life Characteristics Collaborations Communication Complex Computer Models Crystallography Data Development Distal Distant Educational process of instructing Engineering Environment Epitopes Evaluation Family Goals Hand Human Institutes Invaded Investigation Laboratories Lead Libraries Link Malignant Neoplasms Mammalian Cell Mediating Mentors Metastatic to Methods Modeling Modernization Molecular Mutation Neoplasm Metastasis Pathway interactions Peptides Permeability Primary Neoplasm Process Proline Property Protein Engineering Protein Family Protein Isoforms Proteins Proteome RNA Splicing Regulation Regulator Genes Research Research Methodology Research Training Role Site Specificity Structure Techniques Testing Therapeutic Tissues Training Transfection Travel Variant X-Ray Crystallography anticancer research base biophysical techniques cancer cell cancer invasiveness cell motility design experimental study genetic regulatory protein high throughput screening inhibitor insight knock-down member metastatic process neoplastic cell overexpression paralogous gene preference protein function protein protein interaction responsible research conduct scaffold screening skills small molecule inhibitor therapeutic evaluation tool tumor vasodilator-stimulated phosphoprotein

项目摘要

Project Summary/Abstract Cancer metastasis depends on coordinated cytoskeletal processes induced by a characteristic change in expression of specific motility and actin-regulatory genes. Mena, a member of the Ena/VASP family of actin regulatory proteins, is highly upregulated in invasive cancer cells. The Ena/VASP proteins localize to actin-based assemblies via their structurally similar EVH1 domains which bind to short linear motifs (SLMs) in other proteins. Mena is integral to motility pathways that are characteristic of invasive cancer cells. An invasion-associated splice variant of Mena, MenaINV, has far more potent effects on metastasis than Mena and is preferentially expressed in invasive cancer cells. However, determination of the precise mechanistic roles of Mena and MenaINV in metastatic processes has proven challenging. There is currently no molecular explanation for differences in the protein-protein interaction properties of Mena and its paralogs/isoforms. Designed peptide/mini-protein binders can reveal molecular determinants of binding specificity and inspire/inform the design of lead inhibitors. They can also be used to probe the function of proteins in their cellular context and with temporal control, providing a direct evaluation of therapeutic potential. The primary goal of this proposal is to uncover the molecular basis for differences in the protein-interaction properties of Mena and MenaINV, determine the molecular origin of the binding specificity of an existing mini-protein inhibitor, and use this information to design and test paralog- and isoform- selective, cell-permeable mini-protein inhibitors of Mena. This goal will be accomplished by applying an array of biophysical experiments such as NMR, SAXS, X-ray crystallography and binding assays to uncover the molecular origin of protein-interaction differences between Mena and its paralogs/isoforms. These experiments will reveal molecular determinants of inhibitor binding specificity and contribute to our understanding of metastasis by providing a molecular explanation for differences between Mena and MenaINV. The biophysical information gained from these experiments will then be incorporated into the design of paralog- and isoform- specific mini-protein inhibitors using cutting-edge protein design methods, including structure-based computation, focused library design, and high throughput screening techniques. In addition to training in a variety of new research methods, the training plan outlined here includes extensive development of scientific communication, responsible conduct of research, scientific networking, teaching, mentoring and management skills. The research and training will take place in the laboratory of Dr. Amy Keating, a highly interdisciplinary and collaborative group at the forefront of the protein-protein interaction and protein design fields. The Keating lab is embedded in the MIT Biology department and part of the Koch Institute for Integrative Cancer Research, providing an ideal environment for training, collaboration, and research.

项目总结/文摘