scholarly journals Unification and extensive diversification of M/ORF3-related ion channel proteins in coronaviruses and other nidoviruses

2021 ◽  
Author(s):  
Yongjun Tan ◽  
Theresa Schneider ◽  
Prakash K Shukla ◽  
Mahesh B Chandrasekharan ◽  
L Aravind ◽  
...  
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
M Protein ◽  
Detection Methods ◽  
Homology Detection ◽  
Virion Assembly ◽  
Wide Range ◽  
M Proteins ◽  
Slow Evolution ◽  
Membrane Budding

Abstract The coronavirus, SARS-CoV-2, responsible for the ongoing COVID-19 pandemic, has emphasized the need for a better understanding of the evolution of virus-host interactions. ORF3a in both SARS-CoV-1 and SARS-CoV-2 are ion channels (viroporins) implicated in virion assembly and membrane budding. Using sensitive profile-based homology detection methods, we unify the SARS-CoV ORF3a family with several families of viral proteins, including ORF5 from MERS-CoVs, proteins from beta-CoVs (ORF3c), alpha-CoVs (ORF3b), most importantly, the Matrix (M) proteins from CoVs, and more distant homologs from other nidoviruses. We present computational evidence that these viral families might utilize specific conserved polar residues to constitute an aqueous pore within the membrane-spanning region. We reconstruct an evolutionary history of these families and objectively establish the common origin of the M proteins of CoVs and Toroviruses. We also show that the divergent ORF3 clade (ORF3a/ORF3b/ORF3c/ORF5 families) represents a duplication stemming from the M protein in alpha- and beta-CoVs. By phyletic profiling of major structural components of primary nidoviruses, we present a hypothesis for their role in virion assembly of CoVs, ToroVs and Arteriviruses. The unification of diverse M/ORF3 ion channel families in a wide range of nidoviruses, especially the typical M protein in CoVs, reveal a conserved, previously under-appreciated role of ion channels in virion assembly and membrane budding. We show that M and ORF3 are under different evolutionary pressures; in contrast to the slow evolution of M as core structural component, the ORF3 clade is under selection for diversification, which suggests it might act at the interface with host molecules and/or immune attack.

scholarly journals Unification of the M/ORF3-related proteins points to a diversified role for ion conductance in pathogenesis of coronaviruses and other nidoviruses

2020 ◽  
Author(s):  
Yongjun Tan ◽  
Theresa Schneider ◽  
Prakash K. Shukla ◽  
Mahesh B. Chandrasekharan ◽  
L Aravind ◽  
...  
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
M Protein ◽  
Homology Detection ◽  
Virion Assembly ◽  
Host Molecules ◽  
Wide Range ◽  
M Proteins ◽  
Immune Attack ◽  
Selection For

ABSTRACTThe new coronavirus, SARS-CoV-2, responsible for the COVID-19 pandemic has emphasized the need for a better understanding of the evolution of virus-host conflicts. ORF3a in both SARS-CoV-1 and SARS-CoV-2 are ion channels (viroporins) and involved in virion assembly and membrane budding. Using sensitive profile-based homology detection methods, we unify the SARS-CoV ORF3a family with several families of viral proteins, including ORF5 from MERS-CoVs, proteins from beta-CoVs (ORF3c), alpha-CoVs (ORF3b), most importantly, the Matrix (M) proteins from CoVs, and more distant homologs from other nidoviruses. By sequence analysis and structural modeling, we show that these viral families utilize specific conserved polar residues to constitute an ion-conducting pore in the membrane. We reconstruct the evolutionary history of these families, objectively establish the common origin of the M proteins of CoVs and Toroviruses. We show that the divergent ORF3a/ORF3b/ORF5 families represent a duplication stemming from the M protein in alpha- and beta-CoVs. By phyletic profiling of major structural components of primary nidoviruses, we present a model for their role in virion assembly of CoVs, ToroVs and Arteriviruses. The unification of diverse M/ORF3 ion channel families in a wide range of nidoviruses, especially the typical M protein in CoVs, reveal a conserved, previously under-appreciated role of ion channels in virion assembly, membrane fusion and budding. We show that the M and ORF3 are under differential evolutionary pressures; in contrast to the slow evolution of M as core structural component, the CoV-ORF3 clade is under selection for diversification, which indicates it is likely at the interface with host molecules and/or immune attack.IMPORTANCECoronaviruses (CoVs) have become a major threat to human welfare as the causative agents of several severe infectious diseases, namely Severe Acute Respiratory Syndrome (SARS), Middle Eastern Respiratory Syndrome (MERS), and the recently emerging human coronavirus disease 2019 (COVID-19). The rapid spread, severity of these diseases, as well as the potential re-emergence of other CoV-associated diseases have imposed a strong need for a thorough understanding of function and evolution of these CoVs. By utilizing robust domain-centric computational strategies, we have established homologous relationships between many divergent families of CoV proteins, including SARS-CoV/SARS-CoV-2 ORF3a, MERS-CoV ORF5, proteins from both beta-CoVs (ORF3c) and alpha-CoVs (ORF3b), the typical CoV Matrix proteins, and many distant homologs from other nidoviruses. We present evidence that they are active ion channel proteins, and the Cov-specific ORF3 clade proteins are under selection for rapid diversification, suggesting they might have been involved in interfering host molecules and/or immune attack.

scholarly journals Computational Ion Channel Research: from the Application of Artificial Intelligence to Molecular Dynamics Simulations.

2020 ◽  
Vol 55 (S3) ◽  
pp. 14-45
Keyword(s):  
Artificial Intelligence ◽  
Molecular Dynamics ◽  
Ion Channels ◽  
Ion Channel ◽  
Computational Methods ◽  
Homology Modelling ◽  
Channel Structure ◽  
Learning Approaches ◽  
Qsar Analysis ◽  
Wide Range

Although ion channels are crucial in many physiological processes and constitute an important class of drug targets, much is still unclear about their function and possible malfunctions that lead to diseases. In recent years, computational methods have evolved into important and invaluable approaches for studying ion channels and their functions. This is mainly due to their demanding mechanism of action where a static picture of an ion channel structure is often insufficient to fully understand the underlying mechanism. Therefore, the use of computational methods is as important as chemical-biological based experimental methods for a better understanding of ion channels. This review provides an overview on a variety of computational methods and software specific to the field of ion-channels. Artificial intelligence (or more precisely machine learning) approaches are applied for the sequence-based prediction of ion channel family, or topology of the transmembrane region. In case sufficient data on ion channel modulators is available, these methods can also be applied for quantitative structureactivity relationship (QSAR) analysis. Molecular dynamics (MD) simulations combined with computational molecular design methods such as docking can be used for analysing the function of ion channels including ion conductance, different conformational states, binding sites and ligand interactions, and the influence of mutations on their function. In the absence of a three-dimensional protein structure, homology modelling can be applied to create a model of your ion channel structure of interest. Besides highlighting a wide range of successful applications, we will also provide a basic introduction to the most important computational methods and discuss best practices to get a rough idea of possible applications and risks.

Molecular assembly of measles and Nipah virus: specific lipid binding drives conformational change and matrix polymerization

2021 ◽  
Author(s):  
Michael J. Norris ◽  
Monica L. Husby ◽  
William B. Kiosses ◽  
Jieyun Yin ◽  
Linda J. Rennick ◽  
...  
Keyword(s):  
Measles Virus ◽  
Membrane Lipids ◽  
Nipah Virus ◽  
Disease Outbreaks ◽  
Molecular Assembly ◽  
Lipid Binding ◽  
M Protein ◽  
Clear Understanding ◽  
Virion Assembly ◽  
M Proteins

Measles virus, Nipah virus, and multiple other paramyxoviruses cause disease outbreaks in humans and animals worldwide. The paramyxovirus matrix (M) protein mediates virion assembly and budding from host cell membranes. M is thus a key target for antivirals, but few high-resolution structures of paramyxovirus M are available, and we lack the clear understanding of how viral M proteins interact with membrane lipids to mediate viral assembly and egress needed to guide antiviral design. Here, we reveal that M proteins associate with phosphatidylserine and phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) at the plasma membrane. Using X-ray crystallography, electron microscopy, and molecular dynamics we demonstrate that PI(4,5)P2 binding induces conformational and electrostatic changes in the M protein surface that trigger membrane deformation, matrix layer polymerization, and virion assembly.

scholarly journals Naked mole-rat acid-sensing ion channel 3 forms nonfunctional homomers, but functional heteromers

2017 ◽  
Author(s):  
Laura-Nadine Schuhmacher ◽  
Gerard Callejo ◽  
Shyam Srivats ◽  
Ewan St. John Smith
Keyword(s):  
Carbon Dioxide ◽  
Ion Channels ◽  
Ion Channel ◽  
Drg Neurons ◽  
Whole Cell Patch Clamp ◽  
Mole Rat ◽  
Wide Range ◽  
Naked Mole Rat ◽  
Patch Clamp Electrophysiology ◽  
Carbon Dioxide Levels

ABSTRACTAcid-sensing ion channels (ASICs) form both homotrimeric and heterotrimeric ion channels that are activated by extracellular protons and are involved in a wide range of physiological and pathophysiological processes, including pain and anxiety. ASIC proteins can form both homotrimeric and heterotrimeric ion channels. The ASIC3 subunit has been shown to be of particular importance in the peripheral nervous system with pharmacological and genetic manipulations demonstrating a role in pain. Naked mole-rats, despite having functional ASICs, are insensitive to acid as a noxious stimulus and show diminished avoidance of acidic fumes, ammonia and carbon dioxide. Here we cloned naked mole-rat ASIC3 (nmrASIC3) and used a cell surface biotinylation assay to demonstrate that it traffics to the plasma membrane, but using whole-cell patch-clamp electrophysiology we observed that nmrASIC3 is insensitive to both protons and the non-proton ASIC3 agonist 2-Guanidine-4-methylquinazoline (GMQ). However, in line with previous reports of ASIC3 mRNA expression in dorsal root ganglia (DRG) neurons, we found that the ASIC3 antagonist APETx2 reversibly inhibits ASIC-like currents in naked mole-rat DRG neurons. We further show that like the proton-insensitive ASIC2b and ASIC4, nmrASIC3 forms functional, proton sensitive heteromers with other ASIC subunits. An amino acid alignment of ASIC3s between 9 relevant rodent species and human identified unique sequence differences that might underlie the proton insensitivity of nmrASIC3. However, introducing nmrASIC3 differences into rat ASIC3 (rASIC3) produced only minor differences in channel function, and replacing nmrASIC3 sequence with that of rASIC3 did not produce a proton-sensitive ion channel. Our observation that nmrASIC3 forms nonfunctional homomers may reflect a further adaptation of the naked mole-rat to living in an environment with high-carbon dioxide levels.

scholarly journals Briefing in Application of Machine Learning Methods in Ion Channel Prediction

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Hao Lin ◽  
Wei Chen
Keyword(s):  
Machine Learning ◽  
Ion Channels ◽  
Ion Channel ◽  
Inorganic Ions ◽  
Correct Identification ◽  
Biological Processes ◽  
Learning Methods ◽  
Channel Prediction ◽  
Machine Learning Methods ◽  
Wide Range

In cells, ion channels are one of the most important classes of membrane proteins which allow inorganic ions to move across the membrane. A wide range of biological processes are involved and regulated by the opening and closing of ion channels. Ion channels can be classified into numerous classes and different types of ion channels exhibit different functions. Thus, the correct identification of ion channels and their types using computational methods will provide in-depth insights into their function in various biological processes. In this review, we will briefly introduce and discuss the recent progress in ion channel prediction using machine learning methods.

Modelling activity-dependent changes of velocity in C-fibers

2017 ◽  
Vol 16 (1) ◽  
pp. 186-187
Author(s):  
J. Tigerholm ◽  
M.E. Petersson ◽  
O. Obreja ◽  
A. Lampert ◽  
R. Carr ◽  
...  
Keyword(s):  
Neuropathic Pain ◽  
Ion Channels ◽  
Ion Channel ◽  
Small Diameter ◽  
Intracellular Sodium ◽  
Low Frequencies ◽  
C Fibers ◽  
Wide Range ◽  
Activity Dependent ◽  
Insight Into

AbstractAimsWhen C-nociceptors are activated repeatedly using electrical stimulation at relatively low frequencies (0.125–2 Hz), their propagation velocity will decrease. This is referred to as activity-dependent slowing (ADS). The main reason why activity-dependent changes in velocity are of interest is that they can be recorded directly, using invasive methods (microneurography), in patients with chronic pain. Interestingly, in certain patients with neuropathic pain, reduced activity-dependent slowing of conduction has been observed, indicating that these axons have an increased excitability. Through a computational model, it is possible to link such velocity alterations with changes in active conductances, opening for an understanding the underlying excitability changes occurring in these patients.MethodsWe have developed a detailed multicompartment model of a C-nociceptor fiber. This model incorporates a wide range of voltage-gated ion channels (Nav1.7, Nav1.8, Nav1.9, Kdr, KA, KM, KNa and h) which were implemented across a detailed and realistic axon morphology.ResultsThe model predicts that the small diameter of the axon can accumulate intracellular sodium when it is repeatedly activated in a similar fashion as during single fiber microneurography. This increase of intracellular sodium concentration can shift the balance between ion channel currents, shift the membrane potential and membrane input resistance, and thereby generate activity-dependent changes of velocity, such as ADS as well as recovery cycle supernormality.ConclusionsOur results thus provide insight into how activity-dependent excitability changes can be generated in C-fibers. By identifying which ion channels are contributing to activity-dependent changes of velocity this could provide insight into ion channel alterations in neuropathic pain patients.

scholarly journals Detectivity optimization to detect of ultraweak light fluxes with an EM-CCD as binary photon counter array

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ibtissame Khaoua ◽  
Guillaume Graciani ◽  
Andrey Kim ◽  
François Amblard
Keyword(s):  
Dynamic Range ◽  
Signal To Noise Ratio ◽  
Photon Counting ◽  
Detection Methods ◽  
Strong Nonlinearity ◽  
Wide Range ◽  
Depth Analysis ◽  
Spatially Extended ◽  
Extended Sources ◽  
Photon Counting Mode

AbstractFor a wide range of purposes, one faces the challenge to detect light from extremely faint and spatially extended sources. In such cases, detector noises dominate over the photon noise of the source, and quantum detectors in photon counting mode are generally the best option. Here, we combine a statistical model with an in-depth analysis of detector noises and calibration experiments, and we show that visible light can be detected with an electron-multiplying charge-coupled devices (EM-CCD) with a signal-to-noise ratio (SNR) of 3 for fluxes less than $$30\,{\text{photon}}\,{\text{s}}^{ - 1} \,{\text{cm}}^{ - 2}$$ 30 photon s - 1 cm - 2 . For green photons, this corresponds to 12 aW $${\text{cm}}^{ - 2}$$ cm - 2 ≈ $$9{ } \times 10^{ - 11}$$ 9 × 10 - 11 lux, i.e. 15 orders of magnitude less than typical daylight. The strong nonlinearity of the SNR with the sampling time leads to a dynamic range of detection of 4 orders of magnitude. To detect possibly varying light fluxes, we operate in conditions of maximal detectivity $${\mathcal{D}}$$ D rather than maximal SNR. Given the quantum efficiency $$QE\left( \lambda \right)$$ Q E λ of the detector, we find $${ \mathcal{D}} = 0.015\,{\text{photon}}^{ - 1} \,{\text{s}}^{1/2} \,{\text{cm}}$$ D = 0.015 photon - 1 s 1 / 2 cm , and a non-negligible sensitivity to blackbody radiation for T > 50 °C. This work should help design highly sensitive luminescence detection methods and develop experiments to explore dynamic phenomena involving ultra-weak luminescence in biology, chemistry, and material sciences.

scholarly journals Defects and uncertainties of adhesively bonded composite joints

2021 ◽  
Vol 3 (9) ◽  
Author(s):  
Sadik Omairey ◽  
Nithin Jayasree ◽  
Mihalis Kazilas
Keyword(s):  
Composite Materials ◽  
Composite Structures ◽  
Production Efficiency ◽  
Adhesive Bonding ◽  
Detection Methods ◽  
Bonded Joints ◽  
Adhesively Bonded ◽  
Adhesively Bonded Joints ◽  
Wide Range ◽  
Bonded Composite

AbstractThe increasing use of fibre reinforced polymer composite materials in a wide range of applications increases the use of similar and dissimilar joints. Traditional joining methods such as welding, mechanical fastening and riveting are challenging in composites due to their material properties, heterogeneous nature, and layup configuration. Adhesive bonding allows flexibility in materials selection and offers improved production efficiency from product design and manufacture to final assembly, enabling cost reduction. However, the performance of adhesively bonded composite structures cannot be fully verified by inspection and testing due to the unforeseen nature of defects and manufacturing uncertainties presented in this joining method. These uncertainties can manifest as kissing bonds, porosity and voids in the adhesive. As a result, the use of adhesively bonded joints is often constrained by conservative certification requirements, limiting the potential of composite materials in weight reduction, cost-saving, and performance. There is a need to identify these uncertainties and understand their effect when designing these adhesively bonded joints. This article aims to report and categorise these uncertainties, offering the reader a reliable and inclusive source to conduct further research, such as the development of probabilistic reliability-based design optimisation, sensitivity analysis, defect detection methods and process development.

scholarly journals Role of Small Envelope Protein in Sustaining the Intracellular and Extracellular Levels of Hepatitis B Virus Large and Middle Envelope Proteins

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 613
Author(s):  
Jing Zhang ◽  
Yongxiang Wang ◽  
Shuwen Fu ◽  
Quan Yuan ◽  
Qianru Wang ◽  
...  
Keyword(s):  
Envelope Proteins ◽  
Full Length ◽  
M Protein ◽  
Intracellular Level ◽  
S Protein ◽  
L Protein ◽  
M Proteins ◽  
B Virus ◽  
Subviral Particle ◽  
Genotype A

Hepatitis B virus (HBV) expresses co-terminal large (L), middle (M), and small (S) envelope proteins. S protein drives virion and subviral particle secretion, whereas L protein inhibits subviral particle secretion but coordinates virion morphogenesis. We previously found that preventing S protein expression from a subgenomic construct eliminated M protein. The present study further examined impact of S protein on L and M proteins. Mutations were introduced to subgenomic construct of genotype A or 1.1mer replication construct of genotype A or D, and viral proteins were analyzed from transfected Huh7 cells. Mutating S gene ATG to prevent expression of full-length S protein eliminated M protein, reduced intracellular level of L protein despite its blocked secretion, and generated a truncated S protein through translation initiation from a downstream ATG. Truncated S protein was secretion deficient and could inhibit secretion of L, M, S proteins from wild-type constructs. Providing full-length S protein in trans rescued L protein secretion and increased its intracellular level from mutants of lost S gene ATG. Lost core protein expression reduced all the three envelope proteins. In conclusion, full-length S protein could sustain intracellular and extracellular L and M proteins, while truncated S protein could block subviral particle secretion.

scholarly journals Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Sensors ◽  
2021 ◽  
Vol 21 (7) ◽  
pp. 2499
Author(s):  
Michael Dillon ◽  
Maja A. Zaczek-Moczydlowska ◽  
Christine Edwards ◽  
Andrew D. Turner ◽  
Peter I. Miller ◽  
...  
Keyword(s):  
Method Development ◽  
Regulatory Control ◽  
Detection Methods ◽  
Control Individual ◽  
Mouse Bioassay ◽  
Future Market ◽  
Site Testing ◽  
Wide Range ◽  
Development And Validation ◽  
Analytical Standards

In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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