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Arf Functional Landscapes

Arf功能景观

基本信息

批准号：
10620733
负责人：
CATHERINE A ROYER
金额：
$ 34.95万
依托单位：
RENSSELAER POLYTECHNIC INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10620733
关键词：
ADP-Ribosylation Factors Alzheimer&apos s Disease Autoimmune Diseases Bacteria Binding Biophysics Brefeldin A Cell Shape Cell membrane Cell physiology Cells Cerebral cortex Chemicals Communicable Diseases Cytokinesis Data Development Disease Endosomes Enzymes Eukaryotic Cell Exclusion Exhibits Fluorescence Free Energy GTP Binding GTPase-Activating Proteins Genetic Goals Golgi Apparatus Grain Guanine Nucleotide Exchange Factors Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Hepatitis C virus Human Infection Invaded Investigation Link Lipids Malignant Neoplasms Maps Membrane Mental Retardation Mitochondria Modeling Modification Molecular Molecular Conformation Monomeric GTP-Binding Proteins Multidimensional NMR Techniques Mutation Mutation Analysis N-terminal Nucleotides Organelles Parkinson Disease Pathway interactions Pharmaceutical Preparations Population Positioning Attribute Protein Conformation Proteins Regulation of Cell Size Salmonella Signal Pathway Signal Transduction Site Solvents Specificity Structure System Therapeutic Virus Yeasts biophysical properties biophysical tools lipid transport malformation molecular dynamics molecular recognition myristoylation nervous system disorder novel pathogenic bacteria pathogenic virus pressure protein structure three dimensional structure tool trafficking vesicle transport

项目摘要

Summary Membrane organization in eukaryotic cells is controlled by ADP ribosylation factors (Arfs), small GTPases that function as molecular switches to activate signaling cascades. Arfs regulate vesicular transport of lipids and proteins between the ER and the Golgi (Class I-Arf1) and endosome-plasma membrane trafficking (Class III-Arf6), implicating Arf function in cytokinesis, cell shape, organelle transport, mitochondrial and lipid droplet function and pH-dependent regulation of cell size. Mutations in Arfs or their partners have been linked to genetic neurological diseases causing severe malformation of the cerebral cortex or mental retardation. Moreover, many pathogenic bacteria and viruses commandeer Arfs as they invade cells, thereby promoting infection. Our overall goal is to understand the nucleotide exchange transitions of Arf GTPases, the mechanisms of which cannot be deduced from their static structures. We hypothesize that the Arf conformations specifically recognized by their cognate exchange factors correspond to significantly disrupted excited states that are populated at very low levels under standard conditions. Specifically, we aim to map the GDP/GTP switches of Arf1 and Arf6 (Aims 1 and 2), and using mutational analysis, establish the underlying molecular mechanisms of their functional specificity (Aim 3). Our approach combines experimental biophysical tools (multi- dimensional NMR, SAXS and fluorescence) with pressure perturbation and coarse-grained molecular dynamics simulations constrained by our data, to provide structural ensembles and pseudo-free energy landscapes that will reveal functionally relevant excited states implicated in Arf function and specificity. These excited state structures will provide novel target sites for inhibiting Arf signaling pathways, offering new avenues for developing approaches to mitigate the invasive capacity of bacteria and viruses. More generally, the pressure-based mapping approach proposed here represents a powerful means to characterize elusive states of proteins implicated in their functions.

总结