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

Mapping, modeling and manipulating the interactions of protein domains that bind short linear motifs

映射、建模和操纵结合短线性基序的蛋白质结构域的相互作用

基本信息

批准号：
9575778
负责人：
AMY E KEATING
金额：
$ 32.5万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9575778
关键词：
Actins Affect Affinity Benchmarking Binding Binding Proteins Biochemical Biological Biology Biophysics Cancer Control Cell surface Cells Cellular biology Characteristics Code Complex Computing Methodologies Consensus Coupled Cytoskeleton Data Data Quality Development Disease Disseminated Malignant Neoplasm Epitopes Family Family member Fiber Filopodia Genetic Translation Geometry Goals Human Lead Libraries Malignant Neoplasms Measurement Measures Mediating Membrane Proteins Methods Modeling Modernization Molecular Biology Molecular Probes Multiprotein Complexes NMR Spectroscopy Neoplasm Metastasis Neurons Peptide Library Peptides Play Proline Protein Binding Domain Protein Family Protein Isoforms Proteins Proteome RNA Interference Reading Reagent Regulation Reporting Research Resistance Resources Role Screening Result Signal Pathway Signal Transduction Specificity Structural Biologist Structural Models Structure Surface Techniques Tertiary Protein Structure Testing Therapeutic Variant Work X-Ray Crystallography base cancer cell cancer invasiveness cell motility chemotherapy chromatin modification computerized tools design experimental study flexibility human disease inhibitor/antagonist insight model design molecular recognition neurodevelopment paralogous gene predictive modeling predictive test preference programs protein function protein protein interaction scaffold screening therapeutic target vasodilator-stimulated phosphoprotein

项目摘要

Many protein-protein interactions are mediated by short, linear motifs that bind selectively to modular, conserved protein domains. It is a long-standing goal of modern biology to understand the molecular recognition code for these types of interactions, because this would help elucidate mechanisms of signal transduction and predict cellular localization, targeting, and substrate selectivity of proteins that contain peptide-recognition domains. The ability to re-design domain-peptide interactions would lead to protein- interaction inhibitors useful for research and therapy. Recent advances in protein and peptide library screening and sequencing provide exciting opportunities to measure domain-peptide interactions in high throughput. In parallel with such measurements, computational methods are needed to interpret screening results and build models that can be used for protein interaction prediction and design. This proposal describes an integrated experimental/computational program for measuring, modeling and designing interactions mediated by structurally conserved EVH1 domains that bind to short, proline-rich motifs. The proposed studies focus on Ena/VASP family proteins critical for regulation of the actin cytoskeleton. Three human paralogs, Mena, VASP and EVL, act as scaffolds to assemble complexes at the ends of actin fibers. Ena/VASP proteins are critical for the formation of filopodia and lamellipodia and for regulation of actin- based cell motility; they also have newly discovered roles in neural development. Mena, and particularly its isoform Mena invasive, control cancer cell invasion and promote resistance to chemotherapy; the EVH1 binding activity of Mena is therefore an attractive therapeutic target. The peptide binding determinants for EVH1 domains are poorly understood. Short consensus binding motifs have been identified in earlier work but are not sufficient to predict which sequences will bind, or with what paralog specificity. Sequence that flanks known motifs modulates binding to EVH1 domains, but this phenomenon has not been systematically studied. In three specific aims, we propose to (1) experimentally identify Ena/VASP EVH1 domain-binding peptides in the human proteome, determine the sequence requirements for binding, and postulate new biologically relevant interaction partners, (2) apply computational structural modeling and experimental structure determination to understand the mechanisms of binding specificity, including the contributions of core-motif and flanking sequences, and (3) design high affinity and paralog- and/or isoform-selective inhibitors of EVH1 domain interactions. Deeper insight into how EVH1 domains engage their partners will advance our understanding of how these proteins contribute to cell motility, including in invasive cancer cells. Inhibitor design will provide reagents for perturbing Ena/VASP protein functions, including the role of Mena in regulating local mRNA translation in neurons. This work will establish a path for mapping and inhibiting domain-peptide interactions that can be applied to many other peptide-recognition domains.

许多蛋白质相互作用是由短的线性基序介导的，这些基序选择性地结合到模块化的，