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Principal Investigator: Young-wook Jun
Organization: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
Fiscal Year: 2019
Award: $202,851
Funding agency: National Cancer Institute
ABSTRACT
Monitoring dynamic protein-protein interactions (PPIs) in situ has been a topic of interest due to the importance
of PPIs in cell signaling regulation and implications in therapeutic interventions. In this proposal, we introduce a
new PPI method based on nanotechnology-assisted magnetic co-concentration (MCo-C). The key component
of this technology that enables this PPI investigation method is the ability of the nanoprobe-microtweezer
system to magnetically co-concentrate the targeted protein and its interacting partners, enabling significant
amplification of weak and transient PPI signals to be detectable with robust signals. MCo-C can induce signal
amplifications more than 100 times (in vitro; >10 times for live cells) and hence promises to identify weak PPIs
that are not detectable with existing PPI methods. Additionally, this method does not involve any harsh post-
treatments (e.g. cell lysis, chemical membrane crosslinking) and hence allows in situ real-time monitoring of
dynamic PPIs in live cells during the course of cell signaling. To demonstrate the capacity of MCo-C, we will
first carry out PPI assays in in vitro model reaction systems using DNA duplex formation/dissociation. Once the
system is optimized, as initial studies, we propose to investigate PPIs of Notch with potential interaction
partners, including Notch (homodimerization), cis-ligand (e.g. Dll4), and β-catenin. Ultimately, we aim to
provide a platform technology for the systematic investigation of PPIs, accelerating our understanding of the
dynamic regulation of interactome networks in cell signaling.
Terms: <Address><Affinity><After Care><After-Treatment><Aftercare><Alzheimer><Alzheimer Type Dementia><Alzheimer disease><Alzheimer sclerosis><Alzheimer syndrome><Alzheimer's><Alzheimer's Disease><Alzheimers Dementia><Alzheimers disease><Area><Assay><Behavior><Beta Cadherin-Associated Protein><Beta-1 Catenin><Binding><Bioassay><Biologic Assays><Biologic Models><Biological Assay><Biological Models><CUL-2><Cancers><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell membrane><Cell surface><Cells><Chemicals><Clinical Trials><Color><Complement><Complement Proteins><Complex><Coupled><Cytolysis><Cytoplasmic Membrane><Cytosol><DNA><Deoxyribonucleic Acid><Detection><Development><Dimerization><Disease><Disorder><Dissociation><Event><FRET><Fluorescence><Fluorescence Photobleaching Recovery><Fluorescence Recovery After Photobleaching><Fluorescence Resonance Energy Transfer><Future><Förster Resonance Energy Transfer><Goals><Homodimerization><Immune Precipitation><Immunoprecipitation><In Situ><In Vitro><Intracellular Communication and Signaling><Investigation><Kinetics><Label><Length><Ligands><Lipid Bilayers><Location><Lysis><Magnetism><Malignant Neoplasms><Malignant Tumor><Maps><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measurement><Measures><Mechanical Stimulation><Mechanics><Mechanoreceptors><Mediating><Membrane><Methods><Model System><Modeling><Molecular Interaction><Molecular Target><Monitor><Nanotechnology><Noise><Organism><PRO2286><Performance><Plasma Membrane><Primary Senile Degenerative Dementia><Process><Property><Protein Dimerization><Protein Dynamics><Proteins><Reaction><Regulation><Research><Resolution><Screening procedure><Signal Transduction><Signal Transduction Systems><Signaling><Specificity><Structure><System><Techniques><Technology><Testing><Therapeutic><Therapeutic Intervention><Thermodynamic><Thermodynamics><Time><Two Hybrid><Yeast One Hybrid System><Yeast One/Two-Hybrid System><Yeasts><base><beta catenin><biological signal transduction><cross-link><crosslink><dementia of the Alzheimer type><developmental><drug efficacy><experiment><experimental research><experimental study><in vitro Model><innovate><innovation><innovative><interest><intervention therapy><lipid bilayer membrane><living system><magnetic><magnetic field><malignancy><mechanical><melting><membrane structure><nano particle><nano probe><nano tech><nano technology><nano tool><nano-sized particle><nano-technological><nanoparticle><nanoprobe><nanosized particle><nanotech><nanotechnological><nanotool><neoplasm/cancer><notch><notch protein><notch receptors><novel><particle><plasmalemma><post treatment><primary degenerative dementia><programs><protein protein interaction><real time monitoring><realtime monitoring><screening><screening tools><senile dementia of the Alzheimer type><small molecule inhibitor><spatiotemporal><temporal measurement><temporal resolution><time measurement><tool><yeast two hybrid system><β-catenin>