LIVIVO - The Search Portal for Life Sciences

zur deutschen Oberfläche wechseln
Advanced search

Search results

Result 1 - 7 of total 7

Search options

  1. Article: High-throughput confocal imaging of quantum dot-conjugated sars-cov-2 spike trimers to track binding and endocytosis in hek293t cells

    Tran, Bruce Nguyen / Oh, Eunkeu / Susumu, Kimihiro / Wolak, Mason / Gorshkov, Kirill

    Journal of visualized experiments. 2022 Apr. 21, , no. 182

    2022  

    Abstract: The development of new technologies for cellular fluorescence microscopy has facilitated high-throughput screening methods for drug discovery. Quantum dots are fluorescent nanoparticles with excellent photophysical properties imbued with bright and ... ...

    Abstract The development of new technologies for cellular fluorescence microscopy has facilitated high-throughput screening methods for drug discovery. Quantum dots are fluorescent nanoparticles with excellent photophysical properties imbued with bright and stable photoluminescence as well as narrow emission bands. Quantum dots are spherical in shape, and with the proper modification of the surface chemistry, can be used to conjugate biomolecules for cellular applications. These optical properties, combined with the ability to functionalize them with biomolecules, make them an excellent tool for investigating receptor-ligand interactions and cellular trafficking. Here, we present a method that uses quantum dots to track the binding and endocytosis of SARS-CoV-2 spike protein. This protocol can be used as a guide for experimentalists looking to utilize quantum dots to study protein-protein interactions and trafficking in the context of cellular physiology.
    Keywords Severe acute respiratory syndrome coronavirus 2 ; biochemical compounds ; drugs ; endocytosis ; fluorescence ; fluorescence microscopy ; ligands ; photoluminescence ; physiology
    Language English
    Dates of publication 2022-0421
    Size p. e63202.
    Publishing place Journal of Visualized Experiments
    Document type Article
    ZDB-ID 2259946-0
    ISSN 1940-087X
    ISSN 1940-087X
    DOI 10.3791/63202
    Database NAL-Catalogue (AGRICOLA)

    More links

    Kategorien

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

  2. Article ; Online: High-throughput Confocal Imaging of Quantum Dot-Conjugated SARS-CoV-2 Spike Trimers to Track Binding and Endocytosis in HEK293T Cells.

    Tran, Bruce Nguyen / Oh, Eunkeu / Susumu, Kimihiro / Wolak, Mason / Gorshkov, Kirill

    Journal of visualized experiments : JoVE

    2022  , Issue 182

    Abstract: The development of new technologies for cellular fluorescence microscopy has facilitated high-throughput screening methods for drug discovery. Quantum dots are fluorescent nanoparticles with excellent photophysical properties imbued with bright and ... ...

    Abstract The development of new technologies for cellular fluorescence microscopy has facilitated high-throughput screening methods for drug discovery. Quantum dots are fluorescent nanoparticles with excellent photophysical properties imbued with bright and stable photoluminescence as well as narrow emission bands. Quantum dots are spherical in shape, and with the proper modification of the surface chemistry, can be used to conjugate biomolecules for cellular applications. These optical properties, combined with the ability to functionalize them with biomolecules, make them an excellent tool for investigating receptor-ligand interactions and cellular trafficking. Here, we present a method that uses quantum dots to track the binding and endocytosis of SARS-CoV-2 spike protein. This protocol can be used as a guide for experimentalists looking to utilize quantum dots to study protein-protein interactions and trafficking in the context of cellular physiology.
    MeSH term(s) Endocytosis ; HEK293 Cells ; Humans ; Quantum Dots ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus/analysis
    Chemical Substances Spike Glycoprotein, Coronavirus ; spike protein, SARS-CoV-2
    Language English
    Publishing date 2022-04-21
    Publishing country United States
    Document type Journal Article ; Video-Audio Media
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/63202
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

  3. Article: A SARS-CoV-2 nucleocapsid protein TR-FRET assay amenable to high-throughput screening.

    Gorshkov, Kirill / Vasquez, Desarey Morales / Chiem, Kevin / Ye, Chengjin / Tran, Bruce Nguyen / de la Torre, Juan Carlos / Moran, Thomas / Chen, Catherine Z / Martinez-Sobrido, Luis / Zheng, Wei

    bioRxiv : the preprint server for biology

    2021  

    Abstract: Drug development for specific antiviral agents against coronavirus disease 2019 (COVID-19) is still an unmet medical need as the pandemic continues to spread globally. Although huge efforts for drug repurposing and compound screens have put forth, only ... ...

    Abstract Drug development for specific antiviral agents against coronavirus disease 2019 (COVID-19) is still an unmet medical need as the pandemic continues to spread globally. Although huge efforts for drug repurposing and compound screens have put forth, only few compounds remain in late stage clinical trials. New approaches and assays are needed to accelerate COVID-19 drug discovery and development. Here we report a time-resolved fluorescence resonance energy transfer-based assay that detects the severe acute respiratory syndrome coronavirus 2 (SARS-CoV‑2) nucleocapsid protein (NP) produced in infected cells. It uses two specific anti-NP monoclonal antibodies (MAbs) conjugated to donor and acceptor fluorophores that produces a robust ratiometric signal for high throughput screening of large compound collections. Using this assay, we measured a half maximal inhibitory concentration (IC
    Language English
    Publishing date 2021-07-05
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2021.07.03.450938
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

  4. Article ; Online: Enrichment of NPC1-deficient cells with the lipid LBPA stimulates autophagy, improves lysosomal function, and reduces cholesterol storage.

    Ilnytska, Olga / Lai, Kimberly / Gorshkov, Kirill / Schultz, Mark L / Tran, Bruce Nguyen / Jeziorek, Maciej / Kunkel, Thaddeus J / Azaria, Ruth D / McLoughlin, Hayley S / Waghalter, Miriam / Xu, Yang / Schlame, Michael / Altan-Bonnet, Nihal / Zheng, Wei / Lieberman, Andrew P / Dobrowolski, Radek / Storch, Judith

    The Journal of biological chemistry

    2021  Volume 297, Issue 1, Page(s) 100813

    Abstract: Niemann-Pick C (NPC) is an autosomal recessive disorder characterized by mutations in the NPC1 or NPC2 genes encoding endolysosomal lipid transport proteins, leading to cholesterol accumulation and autophagy dysfunction. We have previously shown that ... ...

    Abstract Niemann-Pick C (NPC) is an autosomal recessive disorder characterized by mutations in the NPC1 or NPC2 genes encoding endolysosomal lipid transport proteins, leading to cholesterol accumulation and autophagy dysfunction. We have previously shown that enrichment of NPC1-deficient cells with the anionic lipid lysobisphosphatidic acid (LBPA; also called bis(monoacylglycerol)phosphate) via treatment with its precursor phosphatidylglycerol (PG) results in a dramatic decrease in cholesterol storage. However, the mechanisms underlying this reduction are unknown. In the present study, we showed using biochemical and imaging approaches in both NPC1-deficient cellular models and an NPC1 mouse model that PG incubation/LBPA enrichment significantly improved the compromised autophagic flux associated with NPC1 disease, providing a route for NPC1-independent endolysosomal cholesterol mobilization. PG/LBPA enrichment specifically enhanced the late stages of autophagy, and effects were mediated by activation of the lysosomal enzyme acid sphingomyelinase. PG incubation also led to robust and specific increases in LBPA species with polyunsaturated acyl chains, potentially increasing the propensity for membrane fusion events, which are critical for late-stage autophagy progression. Finally, we demonstrated that PG/LBPA treatment efficiently cleared cholesterol and toxic protein aggregates in Purkinje neurons of the NPC1
    MeSH term(s) Animals ; Autophagy/drug effects ; Cholesterol/metabolism ; Endosomes/metabolism ; Fibroblasts/drug effects ; Fibroblasts/metabolism ; HeLa Cells ; Homeostasis/drug effects ; Humans ; Intracellular Signaling Peptides and Proteins/deficiency ; Intracellular Signaling Peptides and Proteins/metabolism ; Lysophospholipids/metabolism ; Lysosomes/drug effects ; Lysosomes/metabolism ; Mice, Inbred C57BL ; Mice, Transgenic ; Models, Biological ; Monoglycerides/metabolism ; Mutation/genetics ; Neural Stem Cells/drug effects ; Neural Stem Cells/metabolism ; Niemann-Pick C1 Protein ; Niemann-Pick Disease, Type C/genetics ; Phosphatidylglycerols/pharmacology ; Purkinje Cells/drug effects ; Purkinje Cells/metabolism ; Sequestosome-1 Protein/metabolism ; Sphingomyelin Phosphodiesterase/metabolism ; Mice
    Chemical Substances Intracellular Signaling Peptides and Proteins ; Lysophospholipids ; Monoglycerides ; NPC1 protein, human ; Niemann-Pick C1 Protein ; Npc1 protein, mouse ; Phosphatidylglycerols ; SQSTM1 protein, human ; Sequestosome-1 Protein ; bis(monoacylglyceryl)phosphate ; Cholesterol (97C5T2UQ7J) ; Sphingomyelin Phosphodiesterase (EC 3.1.4.12)
    Language English
    Publishing date 2021-05-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1016/j.jbc.2021.100813
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    In stock of ZB MED Cologne/Königswinter

    Uc I Zs.108: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
    ab Jg. 2022: Lesesaal (EG)

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

  5. Article: The SARS-CoV-2 cytopathic effect is blocked with autophagy modulators.

    Gorshkov, Kirill / Chen, Catherine Z / Bostwick, Robert / Rasmussen, Lynn / Xu, Miao / Pradhan, Manisha / Tran, Bruce Nguyen / Zhu, Wei / Shamim, Khalida / Huang, Wenwei / Hu, Xin / Shen, Min / Klumpp-Thomas, Carleen / Itkin, Zina / Shinn, Paul / Simeonov, Anton / Michael, Sam / Hall, Matthew D / Lo, Donald C /
    Zheng, Wei

    bioRxiv : the preprint server for biology

    2020  

    Abstract: SARS-CoV-02 is a new type of coronavirus capable of rapid transmission and causing severe clinical symptoms; much of which has unknown biological etiology. It has prompted researchers to rapidly mobilize their efforts towards identifying and developing ... ...

    Abstract SARS-CoV-02 is a new type of coronavirus capable of rapid transmission and causing severe clinical symptoms; much of which has unknown biological etiology. It has prompted researchers to rapidly mobilize their efforts towards identifying and developing anti-viral therapeutics and vaccines. Discovering and understanding the virus' pathways of infection, host-protein interactions, and cytopathic effects will greatly aid in the design of new therapeutics to treat COVID-19. While it is known that chloroquine and hydroxychloroquine, extensively explored as clinical agents for COVID-19, have multiple cellular effects including inhibiting autophagy, there are also dose-limiting toxicities in patients that make clearly establishing their potential mechanisms-of-action problematic. Therefore, we evaluated a range of other autophagy modulators to identify an alternative autophagy-based drug repurposing opportunity. In this work, we found that 6 of these compounds blocked the cytopathic effect of SARS-CoV-2 in Vero-E6 cells with EC
    Keywords covid19
    Language English
    Publishing date 2020-05-28
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2020.05.16.091520
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

  6. Article ; Online: The SARS-CoV-2 Cytopathic Effect Is Blocked by Lysosome Alkalizing Small Molecules.

    Gorshkov, Kirill / Chen, Catherine Z / Bostwick, Robert / Rasmussen, Lynn / Tran, Bruce Nguyen / Cheng, Yu-Shan / Xu, Miao / Pradhan, Manisha / Henderson, Mark / Zhu, Wei / Oh, Eunkeu / Susumu, Kimihiro / Wolak, Mason / Shamim, Khalida / Huang, Wenwei / Hu, Xin / Shen, Min / Klumpp-Thomas, Carleen / Itkin, Zina /
    Shinn, Paul / Carlos de la Torre, Juan / Simeonov, Anton / Michael, Sam G / Hall, Matthew D / Lo, Donald C / Zheng, Wei

    ACS infectious diseases

    2020  Volume 7, Issue 6, Page(s) 1389–1408

    Abstract: Understanding the SARS-CoV-2 virus' pathways of infection, virus-host-protein interactions, and mechanisms of virus-induced cytopathic effects will greatly aid in the discovery and design of new therapeutics to treat COVID-19. Chloroquine and ... ...

    Abstract Understanding the SARS-CoV-2 virus' pathways of infection, virus-host-protein interactions, and mechanisms of virus-induced cytopathic effects will greatly aid in the discovery and design of new therapeutics to treat COVID-19. Chloroquine and hydroxychloroquine, extensively explored as clinical agents for COVID-19, have multiple cellular effects including alkalizing lysosomes and blocking autophagy as well as exhibiting dose-limiting toxicities in patients. Therefore, we evaluated additional lysosomotropic compounds to identify an alternative lysosome-based drug repurposing opportunity. We found that six of these compounds blocked the cytopathic effect of SARS-CoV-2 in Vero E6 cells with half-maximal effective concentration (EC
    MeSH term(s) Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; COVID-19 ; Drug Repositioning ; Humans ; Lysosomes ; SARS-CoV-2
    Chemical Substances Antiviral Agents
    Language English
    Publishing date 2020-12-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ISSN 2373-8227
    ISSN (online) 2373-8227
    DOI 10.1021/acsinfecdis.0c00349
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

  7. Article ; Online: The SARS-CoV-2 cytopathic effect is blocked with autophagy modulators

    Gorshkov, Kirill / Chen, Catherine Z. / Bostwick, Robert / Rasmussen, Lynn / Xu, Miao / Pradhan, Manisha / Tran, Bruce Nguyen / Zhu, Wei / Shamim, Khalida / Huang, Wenwei / Hu, Xin / Shen, Min / Klumpp-Thomas, Carleen / Itkin, Zina / Shinn, Paul / Simeonov, Anton / Michael, Sam / Hall, Matthew D. / Lo, Donald C. /
    Zheng, Wei

    bioRxiv

    Abstract: SARS-CoV-2 is a new type of coronavirus capable of rapid transmission and causing severe clinical symptoms; much of which has unknown biological etiology. It has prompted researchers to rapidly mobilize their efforts towards identifying and developing ... ...

    Abstract SARS-CoV-2 is a new type of coronavirus capable of rapid transmission and causing severe clinical symptoms; much of which has unknown biological etiology. It has prompted researchers to rapidly mobilize their efforts towards identifying and developing anti-viral therapeutics and vaccines. Discovering and understanding the virus9 pathways of infection, host-protein interactions, and cytopathic effects will greatly aid in the design of new therapeutics to treat COVID-19. While it is known that chloroquine and hydroxychloroquine, extensively explored as clinical agents for COVID-19, have multiple cellular effects including inhibiting autophagy, there are also dose-limiting toxicities in patients that make clearly establishing their potential mechanisms-of-action problematic. Therefore, we evaluated a range of other autophagy modulators to identify an alternative autophagy-based drug repurposing opportunity. In this work, we found that 6 of these compounds blocked the cytopathic effect of SARS-CoV-2 in Vero-E6 cells with EC50 values ranging from 2.0 to 13 μM and selectivity indices ranging from 1.5 to >10-fold. Immunofluorescence staining for LC3B and LysoTracker dye staining assays in several cell lines indicated their potency and efficacy for inhibiting autophagy correlated with the measurements in the SARS-CoV-2 cytopathic effect assay. Our data suggest that autophagy pathways could be targeted to combat SARS-CoV-2 infections and become an important component of drug combination therapies to improve the treatment outcomes for COVID-19.
    Keywords covid19
    Language English
    Publishing date 2020-05-21
    Publisher Cold Spring Harbor Laboratory
    Document type Article ; Online
    DOI 10.1101/2020.05.16.091520
    Database COVID19

    Full text online

    Kategorien

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

To top