A5053400296- Bacteriophages and microbial interactions
- Viral Infections and Outbreaks Research
- Viral gastroenteritis research and epidemiology
- SARS-CoV-2 and COVID-19 Research
- Planarian Biology and Electrostimulation
- Hepatitis B Virus Studies
- Ion channel regulation and function
- Cellular transport and secretion
- Photoreceptor and optogenetics research
- Monoclonal and Polyclonal Antibodies Research
- Virus-based gene therapy research
- vaccines and immunoinformatics approaches
- COVID-19 Clinical Research Studies
- Viral Infections and Vectors
- Neurobiology and Insect Physiology Research
- Long-Term Effects of COVID-19
- Advanced Electron Microscopy Techniques and Applications
- Virology and Viral Diseases
- Enzyme Structure and Function
- Retinal Development and Disorders
- Metabolomics and Mass Spectrometry Studies
- Neuroscience and Neural Engineering
- Viral Infectious Diseases and Gene Expression in Insects
- PI3K/AKT/mTOR signaling in cancer
- RNA and protein synthesis mechanisms
La Jolla Institute for Immunology
2020-2024
Montana State University
2010-2024
Pacific Northwest Research Station
2024
Pacific Northwest National Laboratory
2023
Boston University
2022
Indiana University Bloomington
2015-2019
Understanding immune memory to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for improving diagnostics and vaccines assessing the likely future course of COVID-19 pandemic. We analyzed multiple compartments circulating SARS-CoV-2 in 254 samples from 188 cases, including 43 at ≥6 months after infection. Immunoglobulin G (IgG) spike protein was relatively stable over 6+ months. Spike-specific B cells were more abundant 6 than 1 month symptom onset. SARS-CoV-2-specific CD4
Community of antibodies against COVID-19 The severe acute respiratory syndrome coronavirus 2 spike protein is the basis many vaccines and a primary target neutralizing after infection. Coronavirus Immunotherapeutic Consortium (CoVIC), comprising 56 partners across world, has analyzed panel 269 monoclonal (mAbs) and, on competition profiles, sorted 186 mAbs that receptor binding domain into seven communities. Hastie et al . went to structurally analyze representative antibody used pseudovirus...
ABSTRACT Icosahedral viral capsids are obligated to perform a thermodynamic balancing act. Capsids must be stable enough protect the genome until suitable host cell is encountered yet poised bind receptor, initiate entry, navigate cellular milieu, and release their in appropriate replication compartment. In this study, serotypes of adeno-associated virus (AAV), AAV1, AAV2, AAV5, AAV8, were compared with respect physical properties that influence stability. Thermal stability measurements...
ABSTRACT Understanding immune memory to SARS-CoV-2 is critical for improving diagnostics and vaccines, assessing the likely future course of COVID-19 pandemic. We analyzed multiple compartments circulating in 254 samples from 188 cases, including 43 at ≥ 6 months post-infection. IgG Spike protein was relatively stable over 6+ months. Spike-specific B cells were more abundant than 1 month post symptom onset. SARS-CoV-2-specific CD4 + T CD8 declined with a half-life 3-5 By studying antibody,...
Monoclonal antibodies can provide important pre- or post-exposure protection against infectious disease for those not yet vaccinated in individuals that fail to mount a protective immune response after vaccination. Inmazeb (REGN-EB3), three-antibody cocktail Ebola virus, lessened and improved survival controlled trial. Here, we present the cryo-EM structure at 3.1 Å of virus glycoprotein, determined without symmetry averaging, simultaneous complex with cocktail. This allows modeling...
Voltage clamp fluorometry (VCF) is a powerful technique in which the voltage of cell's membrane clamped to control voltage-sensitive proteins while simultaneously measuring fluorescent signals from protein interest. By combining fluorescence measurements with electrophysiology, VCF provides real-time measurement protein's motions, gives insight into its function. This protocol describes use study protein, voltage-sensing phosphatase (VSP). VSP 3 and 5 phosphatidylinositol phosphate (PIP)...
The SARS-CoV-2 spike protein is synthesized in the endoplasmic reticulum of host cells, from where it undergoes export to Golgi and plasma membrane or retrieval reticulum. Elucidating fundamental principles this bidirectional secretion are pivotal understanding virus assembly designing next generation genetic vaccine with enhanced properties. However, widely used strategy C-terminal affinity tagging cytosolic tail interferes proper trafficking. Hence, structural biophysical investigations...
Weak association energy can lead to uniform nanostructures: defects anneal due subunit lability. What happens when strong leads particles where are trapped? Alphaviruses enveloped viruses whose icosahedral nucleocapsid core assemble independently. We used a simplest case system study Ross River virus (RRV) core-like particle (CLP) self-assembly using purified capsid protein and short DNA oligomer. find that binds the oligomer with high affinity form an assembly competent unit (U)....
Abstract The envelope glycoprotein GP of the ebolaviruses is essential for host cell entry and primary target antibody response. heavily glycosylated with up to 17 N-linked sites, numerous O-linked glycans in its disordered mucin-like domain (MLD), three predicted C-linked mannosylation sites. Glycosylation important attachment, stability fusion activity, shielding from neutralization by serum antibodies. Here, we use glycoproteomics profile site-specific glycosylation patterns ebolavirus...
The voltage-sensing domain (VSD) is a four-helix modular protein that converts electrical signals into conformational changes, leading to open pores and active enzymes. In most proteins, the VSDs do not interact with one another, S1–S3 helices are considered mainly scaffolding, except in phosphatase (VSP) proton channel (Hv). To investigate its contribution VSP function, we mutated four hydrophobic amino acids S1 alanine (F127, I131, I134, L137), individually or combination. Most of these...
Sindbis virus (SINV) is an enveloped, single-stranded RNA virus, which transmitted via mosquitos to a wide range of vertebrate hosts. SINV produced by vertebrate, baby hamster kidney (BHK) cells more than order magnitude less infectious from mosquito (C6/36) cells. The cause this difference poorly understood. In study, charge detection mass spectrometry was used determine the masses intact particles isolated BHK and C6/36 measured are substantially different: 52.88 MDa for derived 50.69...
In vitro assembly of alphavirus nucleocapsid cores, called core-like particles (CLPs), requires a polyanionic cargo. There are no sequence or structure requirements to encapsidate single-stranded nucleic acid this work, we wanted determine how the length cargo impacts stability and assembled CLPs. We hypothesized that neutralizes basic region capsid protein if is long enough, it will also act scaffold CP monomers together. Experimentally found CLPs encapsidating short 27mer oligonucleotides...
Multimerization is a key characteristic of most voltage-sensing proteins. The main exception was thought to be the Ciona intestinalis phosphatase (Ci-VSP). In this study, we show that multimerization also critical for Ci-VSP function. Using coimmunoprecipitation and single-molecule pull-down, find stoichiometry flexible. It exists as both monomers dimers, with dimers favored at higher concentrations. We strong dimerization via domain (VSD) weak domain. voltage-clamp fluorometry, VSDs...
Differential Scanning Fluorimetry (DSF) is a rapid, economical, and straightforward technique for estimating the thermal stability of proteins. The principle involves binding fluorescent dye to thermally exposed hydrophobic pockets protein. dyes used in this are highly non-polar environment quenched when aqueous solution. change fluorescence can be follow unfolding proteins induced by temperature, pH, or chaotropic agents. method well characterized monomeric Here, we extend application...
Swelling of the plant virus CCMV has been monitored in real time, revealing that transition occurs rapidly, is readily reversible, and accompanied by a change particle rigidity.
Voltage-sensing phosphatases (VSPs) are transmembrane proteins that couple changes in membrane potential to hydrolysis of inositol signaling lipids. VSPs catalyze the dephosphorylation phosphatidylinositol phosphates (PIPs) regulate diverse aspects cell physiology including division, growth and migration. highly conserved among chordates, their RNA transcripts have been detected adult embryonic stages frogs, fish, chickens, mice humans. However, subcellular localization biological function...
Chemical and sedimentation procedures are used to purify virus particles. While these approaches successful for wild-type viruses, they often not feasible purifying mutant viruses with assembly defects. We combined two published methods ( Atasheva et al., 2013 ; Moller- Tank ), generate a protocol that uses low-speed centrifugation both wildtype enveloped particles at high yield minimal handling steps. This has successfully been alphavirus imaging structural studies Wang 2015 Ramsey 2017 ).
Ebolavirus disease (EVD) is caused by multiple species of
ABSTRACT The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has accumulated more than 700 million infection cases and 6.9 deaths. New variants have affected antibody interaction with surface spike protein. We defined domain specificities measured hexa-proline stabilized protein (HexaPro) binding kinetics of a large panel antibodies sourced Coronavirus Immunotherapeutics Consortium. Epitope binning analysis competing for HexaPro separated fine majority to four regions: top, outer,...
Abstract The envelope glycoprotein GP of the ebolaviruses is essential for host cell attachment and entry. It also primary target protective neutralizing antibody response in both natural infection vaccination. heavily glycosylated with up to 17 predicted N-linked sites, numerous O-linked glycans its disordered mucin-like domain (MLD), three C-linked mannosylation sites. Glycosylation important cell-surface lectins, as well stability fusion activity. Moreover, it has been shown shield from...