A5009330527- HIV Research and Treatment
- HIV/AIDS drug development and treatment
- HIV/AIDS Research and Interventions
- Immune Cell Function and Interaction
- Cytomegalovirus and herpesvirus research
- Cell death mechanisms and regulation
- Protein Degradation and Inhibitors
- Protein Structure and Dynamics
- Mass Spectrometry Techniques and Applications
- Multiple Myeloma Research and Treatments
- Venomous Animal Envenomation and Studies
- Hepatitis C virus research
- RNA and protein synthesis mechanisms
- Autophagy in Disease and Therapy
- Blood groups and transfusion
- Platelet Disorders and Treatments
- Cardiac tumors and thrombi
- T-cell and B-cell Immunology
- Lipid Membrane Structure and Behavior
- vaccines and immunoinformatics approaches
- Viral-associated cancers and disorders
- Eosinophilic Disorders and Syndromes
- Ocular Diseases and Behçet’s Syndrome
- T-cell and Retrovirus Studies
- interferon and immune responses
University of Pittsburgh
2014-2024
Nef is an HIV-1 virulence factor that promotes viral pathogenicity by altering host cell signaling pathways. binds several members of the Src kinase family, and these interactions have been implicated in pathogenesis HIV/AIDS. However, direct effect interaction on family (SFK) regulation activity has not systematically addressed. We explored this issue using Saccharomyces cerevisiae, a well defined model system for study SFK regulation. Previous studies shown ectopic expression c-Src arrests...
Abstract Treatment of cells with chemotherapy drugs activates the intrinsic mitochondrial pathway apoptosis and caspase protease cascade. Recently, lysosomal cathepsin D has been implicated in caused by oxidative stress, inhibition protein kinase C, stimulation TNFR1 Fas death receptors. However, role chemotherapy-induced cell remained largely unexplored. In this report, we show that treatment U937 leukemia drug etoposide (VP-16) results release into cytosol within 4 hours after initiation...
Nef is an HIV-1 accessory protein essential for AIDS progression and attractive target drug discovery. Lack of a catalytic function makes difficult to assay in chemical library screens. We developed high-throughput screening inhibitors by coupling it one its host cell binding partners, the Src-family kinase Hck. Hck activation dependent upon this assay, providing direct readout activity vitro. Using screen, unique diphenylfuropyrimidine was identified as strong inhibitor Nef-dependent...
HIV-1 Nef triggers down-regulation of cell-surface MHC-I by assembling a Src family kinase (SFK)-ZAP-70/Syk-PI3K cascade. Here, we report that chemical disruption the Nef-SFK interaction with small molecule inhibitor 2c blocks assembly multi-kinase complex and represses HIV-1–mediated in primary CD4 + T-cells. did not interfere PACS-2–dependent trafficking required for or AP-1 PACS-1–dependent sequestering internalized MHC-I, suggesting specifically interfered triggering down-regulation....
The immune receptor signaling pathway is used by nonimmune cells, but the molecular adaptations that underlie its functional diversification are not known. Circulating platelets use homologue glycoprotein VI (GPVI) to respond collagen exposed at sites of vessel injury. In contrast cell responses, platelet activation must take place within seconds successfully form thrombi in flowing blood. Here, we show GPVI utilizes a unique intracellular proline-rich domain (PRD) accelerate activation,...
Eradication of the HIV-1 latent reservoir represents current paradigm to developing a cure for AIDS. has evolved multiple mechanisms evade CD8 T cell responses, including Nef–mediated downregulation MHC-I from surface infected cells. Nef transcripts and protein are detectable in samples aviremic donors, suggesting that expression latently HIV-1–infected CD4 cells protects them immune-mediated clearance. Here, we tested 4 small molecule inhibitors an vitro primary latency model measured...
HIV-1 Nef is a viral accessory protein critical for AIDS progression. lacks intrinsic catalytic activity and binds multiple host cell signaling proteins, including Hck other Src-family tyrosine kinases. binding induces constitutive activation that may contribute to HIV pathogenesis by promoting infectivity, replication downregulation of cell-surface MHC-I molecules. In this study, we developed yeast-based phenotypic screen identify small molecules inhibit the Nef-Hck complex.Nef-Hck...
The HIV-1 Nef accessory factor is critical to the viral life cycle in vivo and promotes immune escape of infected cells via downregulation cell-surface MHC-I. Previously, we discovered small molecules that bind directly block many its functions, including enhancement infectivity replication T cell lines. These compounds also restore MHC-I expression HIV-infected CD4 from AIDS patients, enabling recognition killing by autologous cytotoxic lymphocytes (CTLs). In this study, describe synthesis...
HIV-1 Nef is an attractive target for antiretroviral drug discovery because of its role in promoting infectivity, replication, and host immune system avoidance. Here, we applied a screening strategy which recombinant protein was coupled to activation the Src-family tyrosine kinase Hck, enhances life cycle macrophages. stimulates Hck activity
The HIV-1 virulence factor Nef interacts with the macrophage Src-family kinase Hck, resulting in constitutive activation that contributes to viral replication and immune escape. Previous chemical library screens identified diphenylfuranopyrimdine inhibitor DFP-4AB, which selectively inhibits Nef-dependent Hck activity biochemical assays potently blocks HIV vitro. In present study, hydrogen exchange mass spectrometry (HX MS) was used study conformational changes downregulated result from...
In the companion paper to this work, we described development of a new type hydrogen exchange (HX) mass spectrometry (MS) measurement that integrates Langmuir monolayers. With monolayers, lipid packing density can be reproducibly controlled and changed as desired. Analysis HX in proteins may undergo conformational changes function (for example, rearrangements after insertion into layer) are then possible. We previously used neutron reflection characterize just such change myristoylated HIV-1...
Hydrogen exchange mass spectrometry can be used to compare the conformation and dynamics of proteins that are similar in tertiary structure. If relative deuterium levels measured, differences sequence, forward- back-exchange, peptide retention time, protease digestion patterns all complicate data analysis. We illustrate what learned from such sets by analyzing five variants (Consensus G2E, SF2, NL4-3, ELI, LTNP4) HIV-1 Nef protein, both alone when bound human Hck SH3 domain. Regions with...
SUMMARY The HIV-1 Nef accessory factor is critical to the viral life cycle in vivo where it promotes immune escape of HIV-infected cells and persistence. While these features identify as an attractive antiretroviral drug target, lacks enzymatic activity active site, complicating development occupancy-based drugs. Here we describe proteolysis targeting chimeras (PROTACs) for targeted degradation Nef. Nef-binding compounds, based on a previously reported hydroxypyrazole core, were coupled...
In an interview with Samantha Nelson, a scientific editor of <i>Cell Chemical Biology</i>, the first and corresponding authors research article entitled "PROTAC-mediated degradation HIV-1 Nef efficiently restores cell-surface CD4 MHC-I expression blocks replication" share insights on their paper life as scientists.