scholarly journals The challenge of structural heterogeneity in the native mass spectrometry studies of the SARS-CoV-2 spike protein interactions with its host cell-surface receptor

2021 ◽  
Author(s):  
Yang Yang ◽  
Daniil G. Ivanov ◽  
Igor A. Kaltashov
Keyword(s):  
Mass Spectrometry ◽  
Cell Surface ◽  
Host Cell ◽  
Protein Interactions ◽  
Structural Heterogeneity ◽  
Native Mass Spectrometry ◽  
Cell Surface Receptor ◽  
Spike Protein ◽  
Surface Receptor ◽  
Host Cell Surface

The challenge of structural heterogeneity in the native mass spectrometry studies of the SARS-CoV-2 spike protein interactions with its host cell-surface receptor

2021 ◽  
Author(s):  
Yang Yang ◽  
Daniil G. Ivanov ◽  
Igor A. Kaltashov
Keyword(s):  
Mass Spectrometry ◽  
Cell Surface ◽  
Host Cell ◽  
Structural Heterogeneity ◽  
Native Mass Spectrometry ◽  
Cell Surface Receptor ◽  
Spike Protein ◽  
Native Ms ◽  
Surface Receptor ◽  
Host Cell Surface

Native mass spectrometry (MS) enjoyed tremendous success in the past two decades in a wide range of studies aiming at understanding the molecular mechanisms of physiological processes underlying a variety of pathologies and accelerating the drug discovery process. However, the success record of native MS has been surprisingly modest with respect to the most recent challenge facing the biomedical community - the novel coronavirus infection (COVID-19). The major reason for the paucity of successful studies that use native MS to target various aspects of SARS-CoV-2 interaction with its host is the extreme degree of structural heterogeneity of the viral protein playing a key role in the host cell invasion. Indeed, the SARS-CoV-2 spike protein (S-protein) is extensively glycosylated, presenting a formidable challenge for native mass spectrometry (MS) as a means of characterizing its interactions with both the host cell-surface receptor ACE2 and the drug candidates capable of disrupting this interaction. In this work we evaluate the utility of native MS complemented with the experimental methods using gas-phase chemistry (limited charge reduction) to obtain meaningful information on the association of the S1 domain of the S-protein with the ACE2 ectodomain, and the influence of a small synthetic heparinoid on this interaction. Native MS reveals the presence of several different S1 oligomers in solution and allows the stoichiometry of the most prominent S1/ACE2 complexes to be determined. This enables meaningful interpretation of the changes in native MS that are observed upon addition of a small synthetic heparinoid (the pentasaccharide fondaparinux) to the S1/ACE2 solution, confirming that the small polyanion destabilizes the protein/receptor binding.

scholarly journals Heparan Sulfate Facilitates Spike Protein-Mediated SARS-CoV-2 Host Cell Invasion and Contributes to Increased Infection of SARS-CoV-2 G614 Mutant and in Lung Cancer

2021 ◽  
Vol 8 ◽  
Author(s):  
Jingwen Yue ◽  
Weihua Jin ◽  
Hua Yang ◽  
John Faulkner ◽  
Xuehong Song ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Surface ◽  
Heparan Sulfate ◽  
Host Cell ◽  
Host Cells ◽  
Cell Surface Receptor ◽  
Spike Protein ◽  
Effective Prevention ◽  
Heparin Derivatives ◽  
Host Cell Surface

The severe acute respiratory syndrome (SARS)-like coronavirus disease (COVID-19) is caused by SARS-CoV-2 and has been a serious threat to global public health with limited treatment. Cellular heparan sulfate (HS) has been found to bind SARS-CoV-2 spike protein (SV2-S) and co-operate with cell surface receptor angiotensin-converting enzyme 2 (ACE2) to mediate SARS-CoV-2 infection of host cells. In this study, we determined that host cell surface SV2-S binding depends on and correlates with host cell surface HS expression. This binding is required for SARS-Cov-2 virus to infect host cells and can be blocked by heparin lyase, HS antagonist surfen, heparin, and heparin derivatives. The binding of heparin/HS to SV2-S is mainly determined by its overall sulfation with potential, minor contribution of specific SV2-S binding motifs. The higher binding affinity of SV2-S G614 mutant to heparin and upregulated HS expression may be one of the mechanisms underlying the higher infectivity of the SARS-CoV-2 G614 variant and the high vulnerability of lung cancer patients to SARS-CoV-2 infection, respectively. The higher host cell infection by SARS-CoV-2 G614 variant pseudovirus and the increased infection caused by upregulated HS expression both can be effectively blocked by heparin lyase and heparin, and possibly surfen and heparin derivatives too. Our findings support blocking HS-SV2-S interaction may provide one addition to achieve effective prevention and/treatment of COVID-19.

scholarly journals Significance of thiol-disulfide balance in SARS-CoV-2 infection

2021 ◽  
Vol 72 (3) ◽  
pp. 30-36
Author(s):  
Tatjana Simić
Keyword(s):  
Cell Surface ◽  
Host Cell ◽  
Protein Interactions ◽  
Viral Entry ◽  
Molecular Mechanisms ◽  
Alkylating Agents ◽  
Protein S ◽  
Protein Protein Interactions ◽  
Virus Surface ◽  
Host Cell Surface

Studies of the molecular mechanisms regarding interaction of different viruses with receptors on the host cell surface have shown that the viral entry depends on the specific relationship between free thiol (SH) groups and disulfides on the virus surface, as well as the thiol disulfide balance on the host cell surface. The presence of oxidizing compounds or alkylating agents, which disturb the thiol-disulfide balance on the surface of the virus, can also affect its infectious potential. Disturbed thiol-disulfide balance may also influence protein-protein interactions between SARS-CoV-2 protein S and ACE2 receptors of the host cell. This review presents the basic mechanisms of maintaining intracellular and extracellular thiol disulfide balance and previous experimental and clinical evidence in favor of impaired balance in SARS-CoV-2 infection. Besides, the results of the clinical application or experimental analysis of compounds that induce changes in the thiol disulfide balance towards reduction of disulfide bridges in proteins of interest in COVID-19 infection are presented.

scholarly journals Identification of the Cellular Receptor of Clostridium spiroforme Toxin

2012 ◽  
Vol 80 (4) ◽  
pp. 1418-1423 ◽  
Author(s):  
Panagiotis Papatheodorou ◽  
Claudia Wilczek ◽  
Thilo Nölke ◽  
Gregor Guttenberg ◽  
Daniel Hornuss ◽  
...  
Keyword(s):  
Host Cell ◽  
Cell Receptor ◽  
Cell Surface Receptor ◽  
Target Cells ◽  
Content Type ◽  
Recombinant Production ◽  
Host Cell Receptor ◽  
Surface Receptor ◽  
Microscopic Studies ◽  
Host Cell Surface

ABSTRACTClostridium spiroformeproduces the binary actin-ADP-ribosylating toxin CST (C. spiroformetoxin), which has been proposed to be responsible for diarrhea, enterocolitis, and eventually death, especially in rabbits. Here we report on the recombinant production of the enzyme component (CSTa) and the binding component (CSTb) ofC. spiroformetoxin inBacillus megaterium. By using the recombinant toxin components, we show that CST enters target cells via the lipolysis-stimulated lipoprotein receptor (LSR), which has been recently identified as the host cell receptor of the binary toxinsClostridium difficiletransferase (CDT) andClostridium perfringensiota toxin. Microscopic studies revealed that CST, but not the relatedClostridium botulinumC2 toxin, colocalized with LSR during toxin uptake and traffic to endosomal compartments. Our findings indicate that CST shares LSR withC. difficileCDT andC. perfringensiota toxin as a host cell surface receptor.

scholarly journals Calcitonin/Amylin Receptors and Ramps

2001 ◽  
Vol 1 ◽  
pp. 9-9
Author(s):  
Patrick M. Sexton
Keyword(s):  
Cell Surface ◽  
Protein Interactions ◽  
Receptor Expression ◽  
Cell Surface Receptor ◽  
Calcitonin Receptor ◽  
Receptor Function ◽  
Surface Receptor ◽  
Receptor Activity Modifying Proteins ◽  
G Protein Coupled ◽  
Novel Protein

Our understanding of G protein-coupled receptor function has recently expanded to encompass novel protein interactions that underlie both cell surface receptor expression and the exhibited phenotype. The most notable examples are those involving receptor activity modifying proteins (RAMPs). RAMP association with the calcitonin receptor-like receptor (CRLR) traffics this receptor to the cell surface, where individual RAMPs dictate the expression of unique phenotypes.

scholarly journals SARS-COV2 Envelope Protein (E) interacts with the Lysophosphatidic Acid Receptor 1 (LPAR1) from humans

2020 ◽  
Author(s):  
GIRISH NALLUR
Keyword(s):  
Cell Surface ◽  
Cell Lines ◽  
Protein Interactions ◽  
Biological Significance ◽  
Cell Surface Receptor ◽  
Proteomic Screen ◽  
E Protein ◽  
Cell Surface Staining ◽  
Surface Receptor ◽  
Strong Candidate

Abstract A proteomic screen of human proteins interacting with the SARS-COV2 Envelope (E) protein identified LPAR1 as a strong candidate. Physical association of E protein and LPAR1 was confirmed by co-immunoprecipitation and cell surface staining. LPAR1-E protein interaction was confirmed in all eight human cell lines tested. Many additional proteins participating in the E protein interactions network were also enriched from each of the cell lines, some of which were cell type specific. These findings suggest that LPAR1 is likely a cell surface receptor for the E protein, and pave the way for follow-on studies aimed at understanding the biological significance of the interactions in SARS-COV disease, including the signaling mechanisms.

scholarly journals Computational and in vitro experimental analyses of the anti-COVID-19 potential of Mortaparib and MortaparibPlus

2021 ◽  
Vol 41 (10) ◽  
Author(s):  
Vipul Kumar ◽  
Anissa Nofita Sari ◽  
Hazna Noor Meidinna ◽  
Jaspreet Kaur Dhanjal ◽  
Chandru Subramani ◽  
...  
Keyword(s):  
Cell Surface ◽  
Host Cell ◽  
Antiviral Drug ◽  
Receptor Expression ◽  
Control Measures ◽  
Host Cells ◽  
Cell Surface Receptor ◽  
Viral Inhibition ◽  
Host Cell Surface

Abstract Coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has become a global health emergency. Although new vaccines have been generated and being implicated, discovery and application of novel preventive and control measures are warranted. We aimed to identify compounds that may possess the potential to either block the entry of virus to host cells or attenuate its replication upon infection. Using host cell surface receptor expression (angiotensin-converting enzyme 2 (ACE2) and Transmembrane protease serine 2 (TMPRSS2)) analysis as an assay, we earlier screened several synthetic and natural compounds and identified candidates that showed ability to down-regulate their expression. Here, we report experimental and computational analyses of two small molecules, Mortaparib and MortaparibPlus that were initially identified as dual novel inhibitors of mortalin and PARP-1, for their activity against SARS-CoV-2. In silico analyses showed that MortaparibPlus, but not Mortaparib, stably binds into the catalytic pocket of TMPRSS2. In vitro analysis of control and treated cells revealed that MortaparibPlus caused down-regulation of ACE2 and TMPRSS2; Mortaparib did not show any effect. Furthermore, computational analysis on SARS-CoV-2 main protease (Mpro) that also predicted the inhibitory activity of MortaparibPlus. However, cell-based antiviral drug screening assay showed 30–60% viral inhibition in cells treated with non-toxic doses of either MortaparibPlus or Mortaparib. The data suggest that these two closely related compounds possess multimodal anti-COVID-19 activities. Whereas MortaparibPlus works through direct interactions/effects on the host cell surface receptors (ACE2 and TMPRSS2) and the virus protein (Mpro), Mortaparib involves independent mechanisms, elucidation of which warrants further studies.

scholarly journals Computational and in vitro experimental analyses of the Anti-COVID-19 potential of Mortaparib and MortaparibPlus

2021 ◽  
Author(s):  
Vipul Kumar ◽  
Anissa Nofita Sari ◽  
Hazna Noor Meidinna ◽  
Jaspreet Kaur Dhanjal ◽  
Chandru Subramani ◽  
...  
Keyword(s):  
Cell Surface ◽  
Host Cell ◽  
Receptor Expression ◽  
Control Measures ◽  
Host Cells ◽  
Cell Surface Receptor ◽  
Virus Protein ◽  
Viral Inhibition ◽  
Host Cell Surface

COVID-19 pandemic caused by SARS-CoV-2 virus has become a global health emergency. Although new vaccines have been generated and being implicated, discovery and application of novel preventive and control measures are warranted. We aimed to identify compound/s that may possess the potential to either block the entry of virus to host cells or attenuate its replication upon infection. Using host cell surface receptor expression (Angiotensin-converting enzyme 2 (ACE2) and Transmembrane protease serine 2 (TMPRSS2) analysis as an assay, we earlier screened several synthetic and natural compounds and identified candidates that showed ability to downregulate their expression. Here, we report experimental and computational analyses of two small molecules, Mortaparib and MortaparibPlus that were initially identified as dual novel inhibitors of mortalin and PARP-1, for their activity against SARS-CoV-2. In silico analyses showed that MortaparibPlus, but not Mortaparib, stably binds into the catalytic pocket of TMPRSS2. In vitro analysis of control and treated cells revealed that MortaparibPlus caused downregulation of ACE2 and TMPRSS2; Mortaparib did not show any effect. Furthermore, computational analysis on SARS-CoV-2 main protease (Mpro) that also predicted the inhibitory activity of MortaparibPlus.  However, cell based anti-virus drug screening assay showed 30~60% viral inhibition in cells treated with non-toxic doses of either MortaparibPlus or Mortaparib. The data suggests that these two closely related compounds possess multimodal anti-COVID 19 activities. Whereas MortaparibPlus works through direct interactions/effects on the host cell surface receptors (ACE2 and TMPRSS2) and the virus protein (Mpro), Mortaparib involves independent mechanisms, elucidation of which warrants further studies.

Sign in / Sign up

Export Citation Format

Share Document