Understanding the structural mechanism of spontaneous ubiquitin cargo clustering on the cell plasma membrane
了解细胞质膜上自发泛素货物聚集的结构机制
基本信息
- 批准号:10730734
- 负责人:
- 金额:$ 36.5万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2023
- 资助国家:美国
- 起止时间:2023-09-06 至 2026-08-31
- 项目状态:未结题
- 来源:
- 关键词:Atomic Force MicroscopyBindingBinding ProteinsBiomimetic MaterialsCapsid ProteinsCarrier ProteinsCell membraneCell physiologyCellsCholesterolCollaborationsCommunicationComputer softwareCrohn&aposs diseaseCuesDataData AnalysesDependenceDiseaseEngineeringEnvironmentFluorescenceFluorescence MicroscopyGoalsGolgi ApparatusHealthHomeostasisImageInvestigationLearningLengthLifeLipid BilayersLipidsMalignant NeoplasmsMediatingMembraneMembrane MicrodomainsMembrane ProteinsMethodsModelingMolecularMolecular StructureNamesNon-Insulin-Dependent Diabetes MellitusOrganellesOrganismOrganizational ChangeOutcomePharmaceutical PreparationsPhysical condensationPhysiologicalPlasma CellsPlayPolymersProcessProteinsRAD23B geneRecruitment ActivityResearchRoleSamplingScienceScientistSolidSortingStructureStudentsSynapsesSystemTertiary Protein StructureTestingTherapeuticThermodynamicsTimeTransport ProcessUbiquitinUnderserved PopulationUniversitiesUnsaturated FatsVesicleVirus DiseasesWorkdeep learningdesignenvironmental changeflexibilityinsightinstrumentlipid transportmembrane modelmembrane reconstitutionmicroscopic imagingmolecular modelingnervous system disorderneuralprogramsprotein structureprotein transportreceptorreconstitutionrecruitresponseskillssymposiumtraffickingundergraduate studentunderserved students
项目摘要
Abstract
The PI proposes to study the role of spontaneous Ubiquitin (UB) cargo clustering on the plasma membranes in
vesicular trafficking using the model membrane reconstitution. Vesicular trafficking is a process of material
transport by lipid vesicles between membrane bound organelles. The process is one of the very basic cellular
functions and thus is involved in most life processes of higher organisms. However, its molecular mechanism is
still poorly understood. Ubiquitylation, an enzymatic attachment of UB, is involved in many trafficking processes,
but canonically, UB cargo has been considered only as a passive binder to the downstream trafficking proteins.
Polymeric ubiquitylation of cargos and their multivalent interaction with binder proteins are very common in living
cells leading to spontaneous protein condensate domain formation on the cell membranes. These cargo cluster-
ing domains are also known to very closely interact with lipid raft domains. Understanding the spontaneous
clustering of UB cargo as an active spatial organizer of the vesicular trafficking process can advance our current
understanding of the vesicular trafficking mechanism by providing insight into the traditionally overlooked roles
of UB cargo in the process. The main methods of this investigation are fluorescence microscopy (FM) and atomic
force microscopy (AFM). Image data are analyzed by the deep-learning-assisted analysis software. The goals
of this proposal are: (1) Identifying the structural organization of UB cargo clusters created by proteins with
different structures; (2) Understanding the lipid composition dependence of the UB cluster organization. The PI
places an emphasis on structural understanding of the cargo clustering domain formation in this proposal. FM is
used for high throughput imaging of many samples with different protein structures to understand the structural
cues that lead to clustering domain formation. AFM is used to study the structural organization of the clusters at
the molecular level to obtain direct structural snapshots to test hypothetical models. The proposal will enhance
the research environment at Montclair State University by synergistically enabling the use of the high-sensitivity
AFM instrument the university recently obtained. Moreover, the proposal will address the current high demand
of undergraduate students to participate in research while preparing them to advance to graduate programs in
health-related sciences. In collaboration with the GS-LSAMP program, students of underserved population will
be actively recruited. Students are expected to acquire experimental skills, learn data analysis, and present in
conferences to meet and communicates with scientists in the field.
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摘要
项目成果
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