Results for “human domain” — Page 33

DOE OSTI2022

Bivalent recognition of fatty acyl-CoA by a human integral membrane palmitoyltransferase

S-acylation, also known as palmitoylation, is the most abundant form of protein lipidation in humans. This reversible posttranslational modification, which targets thousands of proteins, is catalyzed by 23 members of the DHHC family of integral membrane enzymes. DHHC enzymes use fatty acyl-CoA as the ubiquitous fatty acyl donor and become autoacylated at a catalytic cysteine; this intermediate subsequently transfers the fatty acyl group to a cysteine in the target protein. Protein S-acylation intersects with almost all areas of human physiology, and several DHHC enzymes are considered as possible therapeutic targets against diseases such as cancer. These efforts would greatly benefit from a detailed understanding of the molecular basis for this crucial enzymatic reaction. Here, in this study, we combine X-ray crystallography with all-atom molecular dynamics simulations to elucidate the structure of the precatalytic complex of human DHHC20 in complex with palmitoyl CoA. The resulting structure reveals that the fatty acyl chain inserts into a hydrophobic pocket within the transmembrane spanning region of the protein, whereas the CoA headgroup is recognized by the cytosolic domain through polar and ionic interactions. Biochemical experiments corroborate the predictions from our structural model. We show, using both computational and experimental analyses, that palmitoyl CoA acts as a bivalent ligand where the interaction of the DHHC enzyme with both the fatty acyl chain and the CoA headgroup is important for catalytic chemistry to proceed. This bivalency explains how, in the presence of high concentrations of free CoA under physiological conditions, DHHC enzymes can efficiently use palmitoyl CoA as a substrate for autoacylation.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

The J-elongated conformation of β 2 -glycoprotein I predominates in solution: Implications for our understanding of antiphospholipid syndrome

β 2 -Glycoprotein I (β 2 GPI) is an abundant plasma protein displaying phospholipid-binding properties. Because it binds phospholipids, it is a target of antiphospholipid antibodies (aPLs) in antiphospholipid syndrome (APS), a life-threatening autoimmune thrombotic disease. Indeed, aPLs prefer membrane-bound β 2 GPI to that in solution. β 2 GPI exists in two almost equally populated redox states: oxidized, in which all the disulfide bonds are formed, and reduced, in which one or more disulfide bonds are broken. Furthermore, β 2 GPI can adopt multiple conformations (i.e. J-elongated, S-twisted, and O-circular). While strong evidence indicates that the J-form is the structure bound to aPLs, which conformation exists and predominates in solution remains controversial, and so is the conformational pathway leading to the bound state. Here, we report that human recombinant β 2 GPI purified under native conditions is oxidized. Moreover, under physiological pH and salt concentrations, this oxidized form adopts a J-elongated, flexible conformation, not circular or twisted, in which the N-terminal domain I (DI) and the C-terminal domain V (DV) are exposed to the solvent. Consistent with this model, binding kinetics and mutagenesis experiments revealed that in solution the J-form interacts with negatively charged liposomes and with MBB2, a monoclonal anti-DI antibody that recapitulates most of the features of pathogenic aPLs. We conclude that the preferential binding of aPLs to phospholipid-bound β 2 GPI arises from the ability of its preexisting J-form to accumulate on the membranes, thereby offering an ideal environment for aPL binding. We propose that targeting the J-form of β 2 GPI provides a strategy to block pathogenic aPLs in APS.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2023

Genomic and phenotypic comparison of two variants of multidrug-resistant Salmonella enterica serovar Heidelberg isolated during the 2015–2017 multi-state outbreak in cattle

Salmonella enterica subspecies enterica serovar Heidelberg (Salmonella Heidelberg) has caused several multistate foodborne outbreaks in the United States, largely associated with the consumption of poultry. However, a 2015–2017 multidrug-resistant (MDR) Salmonella Heidelberg outbreak was linked to contact with dairy beef calves. Traceback investigations revealed calves infected with outbreak strains of Salmonella Heidelberg exhibited symptoms of disease frequently followed by death from septicemia. To investigate virulence characteristics of Salmonella Heidelberg as a pathogen in bovine, two variants with distinct pulse-field gel electrophoresis (PFGE) patterns that differed in morbidity and mortality during the multistate outbreak were genotypically and phenotypically characterized and compared. Strain SX 245 with PFGE pattern JF6X01.0523 was identified as a dominant and highly pathogenic variant causing high morbidity and mortality in affected calves, whereas strain SX 244 with PFGE pattern JF6X01.0590 was classified as a low pathogenic variant causing less morbidity and mortality. Comparison of whole-genome sequences determined that SX 245 lacked ~200 genes present in SX 244, including genes associated with the IncI1 plasmid and phages; SX 244 lacked eight genes present in SX 245 including a second YdiV Anti-FlhC(2)FlhD(4) factor, a lysin motif domain containing protein, and a pentapeptide repeat protein. RNA-sequencing revealed fimbriae-related, flagella-related, and chemotaxis genes had increased expression in SX 245 compared to SX 244. Furthermore, SX 245 displayed higher invasion of human and bovine epithelial cells than SX 244. These data suggest that the presence and up-regulation of genes involved in type 1 fimbriae production, flagellar regulation and biogenesis, and chemotaxis may play a role in the increased pathogenicity and host range expansion of the Salmonella Heidelberg isolates involved in the bovine-related outbreak.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Structure and Antigenicity of the Porcine Astrovirus 4 Capsid Spike

Porcine astrovirus 4 (PoAstV4) has been recently associated with respiratory disease in pigs. In order to understand the scope of PoAstV4 infections and to support the development of a vaccine to combat PoAstV4 disease in pigs, we designed and produced a recombinant PoAstV4 capsid spike protein for use as an antigen in serological assays and for potential future use as a vaccine antigen. Structural prediction of the full-length PoAstV4 capsid protein guided the design of the recombinant PoAstV4 capsid spike domain expression plasmid. The recombinant PoAstV4 capsid spike was expressed in Escherichia coli, purified by affinity and size-exclusion chromatography, and its crystal structure was determined at 1.85 Å resolution, enabling structural comparisons to other animal and human astrovirus capsid spike structures. The recombinant PoAstV4 capsid spike protein was also used as an antigen for the successful development of a serological assay to detect PoAstV4 antibodies, demonstrating that the recombinant PoAstV4 capsid spike retains antigenic epitopes found on the native PoAstV4 capsid. These studies lay a foundation for seroprevalence studies and the development of a PoAstV4 vaccine for swine.

Virology↗

NASA NTRS2001

Adjustable Autonomy Testbed

The Adjustable Autonomy Testbed (AAT) is a simulation-based testbed located in the Intelligent Systems Laboratory in the Automation, Robotics and Simulation Division at NASA Johnson Space Center. The purpose of the testbed is to support evaluation and validation of prototypes of adjustable autonomous agent software for control and fault management for complex systems. The AA T project has developed prototype adjustable autonomous agent software and human interfaces for cooperative fault management. This software builds on current autonomous agent technology by altering the architecture, components and interfaces for effective teamwork between autonomous systems and human experts. Autonomous agents include a planner, flexible executive, low level control and deductive model-based fault isolation. Adjustable autonomy is intended to increase the flexibility and effectiveness of fault management with an autonomous system. The test domain for this work is control of advanced life support systems for habitats for planetary exploration. The CONFIG hybrid discrete event simulation environment provides flexible and dynamically reconfigurable models of the behavior of components and fluids in the life support systems. Both discrete event and continuous (discrete time) simulation are supported, and flows and pressures are computed globally. This provides fast dynamic simulations of interacting hardware systems in closed loops that can be reconfigured during operations scenarios, producing complex cascading effects of operations and failures. Current object-oriented model libraries support modeling of fluid systems, and models have been developed of physico-chemical and biological subsystems for processing advanced life support gases. In FY01, water recovery system models will be developed.

Malin, Jane T.↗

NASA NTRS2023

Using Virtual Reality to Envision Deployment of Spacesuit-Compatible Augmented Reality Displays for Lunar Surface Operations

The National Aeronautics and Space Administration (NASA) aims to land crew on the lunar surface to establish a sustainable presence and develop operational concepts for future long-duration missions. New technologies will be necessary to extend planning and execution capabilities for lunar surface activities. NASA’s Joint Augmented Reality Visual Informatics System (Joint AR) is one such technology. Joint AR is a suit-mounted augmented reality (AR) display and computes system which facilitates unprecedented information exchange and data visualization capabilities between mission support operators and suited crew. This paper describes challenges associated with developing AR technology for an envisioned work domain by applying a sociotechnical lens to the iterative testing and development of novel AR technology through virtual reality (VR). A VR testbed was established to simulate a representative lunar surface environment, enabling a series of three human-in-the-loop (HITL) tests evaluating AR navigation interfaces for exploration extravehicular activity (xEVA). Our findings identify several considerations for future Joint AR design and testing efforts, including challenges with data overload, attentional demands, and environment-related perceptual challenges. Trade-offs and potential approaches are discussed to mitigate these challenges and improve future Joint AR testing fidelity.

Jacob Keller↗

NASA NTRSDate not supplied

Using Virtual Reality to Envision Deployment of Spacesuit-Compatible Augmented Reality Displays for Lunar Surface Operations

The National Aeronautics and Space Administration (NASA) aims to land crew on the lunar surface to establish a sustainable presence and develop operational concepts for future long-duration missions. New technologies will be necessary to extend planning and execution capabilities for lunar surface activities. NASA’s Joint Augmented Reality Visual Informatics System (Joint AR) is one such technology. Joint AR is a suit-mounted augmented reality (AR) display and computes system which facilitates unprecedented information exchange and data visualization capabilities between mission support operators and suited crew. This paper describes challenges associated with developing AR technology for an envisioned work domain by applying a sociotechnical lens to the iterative testing and development of novel AR technology through virtual reality (VR). A VR testbed was established to simulate a representative lunar surface environment, enabling a series of three human-in-the-loop (HITL) tests evaluating AR navigation interfaces for exploration extravehicular activity (xEVA). Our findings identify several considerations for future Joint AR design and testing efforts, including challenges with data overload, attentional demands, and environment-related perceptual challenges. Trade-offs and potential approaches are discussed to mitigate these challenges and improve future Joint AR testing fidelity.

Matthew Miller↗

NASA NTRSDate not supplied

Acquisition of and Access to Research Omics Data

Omics data are essential for understanding the myriad and complex effects of space environments on humans. To assure maximum benefit from these kinds of data, the NASA Human Research Program Data Management Plan stipulates that human omics data should be archived within and accessed through the NASA Life Sciences Portal (NLSP). The NLSP has the capability to acquire and provision access to omics (and other kinds of) research results for individual and ad-hoc groups of subjects at the direction of institutional review boards, or other authorizing bodies or individuals, per institutional, program and investigation-specific policies and procedures. However, because some single-subject omics data, like CT scans and other kinds of large, complex biomedical data, could be used to identify heretofore unknown risks to the subject’s health, or, in certain cases, be used to identify a subject, NASA Policy Directive 7170.1 describes various policies regarding the management of and access to “research genetic testing” data, which includes many kinds of omics data. For example, NPD 7170.1 prohibits access to human research genetic data by NASA personnel who make employment decisions for the subjects from whom the data were obtained. To meet the objective of acquiring research omics data for NLSP in compliance with the policies in NPD 7170.1 and other applicable NASA policies, we designed NOMADS (the NLSP Omics Multimodal Acquisition of Data System), a new component that supports the transfer of large research data files, including research genetic testing data, using one of several different transfer mechanisms. The choice of mechanism is made by the submitter of the data, with guiding information from the system, and is likely to often be determined in large part by the nature and source location of the data. For example, for small files where the source data files are not already stored in a cloud storage system, users are likely to prefer to transfer their data to the NLSP via a web browser. Conversely, for large sets of files already organized and stored in a cloud storage system, users may opt for NOMAD’s cloud-to-cloud transfer method. All omics datasets targeted for the NASA Life Sciences Data Archive must pass a variety of quality checks to ensure data integrity and adherence to the standards defined by the LSDA Data Submission Guidelines (DSG) (see https://nlsp.nasa.gov/explore/lsdahome/datasubmit). These include requirements that data are consistent with open standards established by the omics community. Non-compliant data will not be accepted however archivists are available to advise submitters on how to revise data submissions and re-submit until compliance is achieved. Following compliance with the LSDA DSG, omics data next undergo a variety of additional quality checks to ensure the data meet omics community standards. Domain specific Omics data quality control tools and techniques are continually evolving and linked to the advancements in omics assays utilized and thus, the tools and techniques utilized by the LSDA for data quality control and validation will need to be sustained accordingly. All human omics data will be access controlled according to the policies described above, and requiring IRB approval for any additional access grants once the data are acquired (including access for analysis using the NLSP workspace tools).

Omics↗

NASA NTRS2024

Acquisition of and Access to Research Omics Data

Omics data are essential for understanding the myriad and complex effects of space environments on humans. To assure maximum benefit from these kinds of data, the NASA Human Research Program Data Management Plan stipulates that human omics data should be archived within and accessed through the NASA Life Sciences Portal (NLSP). The NLSP has the capability to acquire and provision access to omics (and other kinds of) research results for individual and ad-hoc groups of subjects at the direction of institutional review boards, or other authorizing bodies or individuals, per institutional, program and investigation-specific policies and procedures. However, because some single-subject omics data, like CT scans and other kinds of large, complex biomedical data, could be used to identify heretofore unknown risks to the subject’s health, or, in certain cases, be used to identify a subject, NASA Policy Directive 7170.1 describes various policies regarding the management of and access to “research genetic testing” data, which includes many kinds of omics data. For example, NPD 7170.1 prohibits access to human research genetic data by NASA personnel who make employment decisions for the subjects from whom the data were obtained. To meet the objective of acquiring research omics data for NLSP in compliance with the policies in NPD 7170.1 and other applicable NASA policies, we designed NOMADS (the NLSP Omics Multimodal Acquisition of Data System), a new component that supports the transfer of large research data files, including research genetic testing data, using one of several different transfer mechanisms. The choice of mechanism is made by the submitter of the data, with guiding information from the system, and is likely to often be determined in large part by the nature and source location of the data. For example, for small files where the source data files are not already stored in a cloud storage system, users are likely to prefer to transfer their data to the NLSP via a web browser. Conversely, for large sets of files already organized and stored in a cloud storage system, users may opt for NOMAD’s cloud-to-cloud transfer method. All omics datasets targeted for the NASA Life Sciences Data Archive must pass a variety of quality checks to ensure data integrity and adherence to the standards defined by the LSDA Data Submission Guidelines (DSG) (see https://nlsp.nasa.gov/explore/lsdahome/datasubmit). These include requirements that data are consistent with open standards established by the omics community. Non-compliant data will not be accepted however archivists are available to advise submitters on how to revise data submissions and re-submit until compliance is achieved. Following compliance with the LSDA DSG, omics data next undergo a variety of additional quality checks to ensure the data meet omics community standards. Domain specific Omics data quality control tools and techniques are continually evolving and linked to the advancements in omics assays utilized and thus, the tools and techniques utilized by the LSDA for data quality control and validation will need to be sustained accordingly. All human omics data will be access controlled according to the policies described above, and requiring IRB approval for any additional access grants once the data are acquired (including access for analysis using the NLSP workspace tools).

Omics↗

DOE OSTI2023

Artificial extracellular matrix scaffolds of mobile molecules enhance maturation of human stem cell-derived neurons

Human induced pluripotent stem cell (hiPSC) technologies offer a unique resource for modeling neurological diseases. However, iPSC models are fraught with technical limitations including abnormal aggregation and inefficient maturation of differentiated neurons. These problems are in part due to the absence of synergistic cues of the native extracellular matrix (ECM). We report on the use of three artificial ECMs based on peptide amphiphile (PA) supramolecular nanofibers. All nanofibers display the laminin-derived IKVAV signal on their surface but differ in the nature of their non-bioactive domains. Here, we find that nanofibers with greater intensity of internal supramolecular motion have enhanced bioactivity toward hiPSC-derived motor and cortical neurons. Proteomic, biochemical, and functional assays reveal that highly mobile PA scaffolds caused enhanced β1-integrin pathway activation, reduced aggregation, increased arborization, and matured electrophysiological activity of neurons. Our work highlights the importance of designing biomimetic ECMs to study the development, function, and dysfunction of human neurons.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2024

In vitro enhancement of Zika virus infection by preexisting West Nile virus antibodies in human plasma-derived immunoglobulins revealed after P2 binding site-specific enrichment

ABSTRACT Human immunoglobulin preparations contain a diverse range of polyclonal antibodies that reflect past immune responses against pathogens encountered by the blood donor population. In this study, we examined a panel of intravenous immunoglobulins (IGIVs) manufactured over the past two decades (1998–2020) for their capacity to neutralize or enhance Zika virus (ZIKV) infectionin vitro. These IGIVs were selected specifically based on their production dates in relation to the occurrences of two flavivirus outbreaks in the U.S.: the West Nile virus (WNV) outbreak in 1999 and the ZIKV outbreak in 2015. As demonstrated by enzyme-linked immunosorbent assay (ELISA) experiments, IGIVs made before the ZIKV outbreak already harbored antibodies that bind to various peptides across the envelope protein of ZIKV because of the WNV outbreak. Using phage display, the most dominant binding site was mapped precisely to the P2 peptide between residues 211 and 230 within domain II, where BF1176-56, an anti-ZIKV monoclonal antibody, also binds. When tested in permissive Vero E6 cells for ZIKV neutralization, the IGIVs, even after undergoing rigorous enrichment for P2 binding specificity, failed, as did BF1176-56. Meanwhile, BF1176-56 enhanced ZIKV infection in both FcγRII-expressing K562 cells and human peripheral blood mononuclear cells. However, for enhancement by the IGIVs to be detected in these cells, a substantial increase in their P2 binding specificity was required, thus linking the P2 site with ZIKV enhancementin vitro. Our findings warrant further study of the significance of elevated levels of anti-WNV antibodies in IGIVs, considering that various mechanisms operatingin vivomay modulate ZIKV infection outcomes. IMPORTANCE We investigated the capacity of intravenous immunoglobulins manufactured previously over two decades (1998–2020) to neutralize or enhance Zika virus infectionin vitro. West Nile virus antibodies in IGIVs could not neutralize Zika virus initially; however, once the IGIVs were concentrated further, they enhanced its infection. These findings lay the groundwork for exploring how preexisting WNV antibodies in IGIVs could impact Zika infection, bothin vitroandin vivo. Our observations are historically significant, since we tested a panel of IGIV lots that were carefully selected based on their production dates which covered two major flavivirus outbreaks in the U.S.: the WNV outbreak in 1999 and the ZIKV outbreak in 2015. These findings will facilitate our understanding of the interplay among closely related viral pathogens, particularly from a historical perspective regarding large blood donor populations. They should remain relevant for future outbreaks of emerging flaviviruses that may potentially affect vulnerable populations.

Microbiology↗

NASA NTRS2003

Lysine hydroxylation of collagen in a fibroblast cell culture system

The lysine (Lys) hydroxylation pattern of type I collagen produced by human fibroblasts in culture was analyzed and compared. Fibroblasts were cultured from normal human skin (NSF), keloid (KDF), fetal skin (FDF), and skin tissues of Ehlers-Danlos syndrome type VIA and VIB patients (EDS-VIA and -VIB). The type I collagen alpha chains with or without non-helical telopeptides were purified from the insoluble matrix and analyzed. In comparison with NSFs, KDF and FDF showed significantly higher Lys hydroxylation, particularly in the telopeptide domains of both alpha chains. Both EDS-VIA and -VIB showed markedly lower Lys hydroxylation in the helical domains of both alpha chains whereas that in the telopeptides was comparable with those of NSFs. A similar profile was observed in the tissue sample of the EDS-VIB patient. These results demonstrate that the Lys hydroxylation pattern is domain-specific within the collagen molecule and that this method is useful to characterize the cell phenotypes in normal/pathological connective tissues.

NASA Discipline Cell Biology↗

DOE OSTI2021

Ultra-large-scale ab initio quantum chemical computation of bio-molecular systems: The case of spike protein of SARS-CoV-2 virus

The COVID-19 pandemic poses a severe threat to human health with an unprecedented social and economic disruption. Spike (S) glycoprotein of the SARS-CoV-2 virus is pivotal in understanding the virus anatomy, since it initiates the first contact with the ACE2 receptor in the human cell. We report results of ab initio computation of the spike protein, the largest ab initio quantum chemical computation to date on any bio-molecular system, using a divide and conquer strategy by focusing on individual structural domains. In this approach we divided the S-protein into seven structural domains: N-terminal domain (NTD), receptor binding domain (RBD), subdomain 1 (SD1), subdomain 2 (SD2), fusion peptide (FP), heptad repeat 1 with central helix (HR1-CH) and connector domain (CD). The entire Chain A has 14,488 atoms including the hydrogen atoms but excluding the amino acids with missing coordinates based on the PDB data (ID: 6VSB). The results include structural refinement, ab initio calculation of intramolecular bonding mechanism, 3- dimensional non-local inter-amino acid interaction with implications for the inter-domain interaction. Details of the electronic structure, interatomic bonding, partial charge distribution and the role played by hydrogen bond network are discussed. In the interaction among structural domains, we present new insights for crucial hinge-like movement and fusion process. Extension of such calculation to the interface between the S-protein binding domain and ACE2 receptor can provide a pathway for computational understanding of mutations and the design of therapeutic drugs to combat the COVID-19 pandemic.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1987

Expert systems for space power supply - Design, analysis, and evaluation

The feasibility of applying expert systems to the conceptual design, analysis, and evaluation of space power supplies in particular, and complex systems in general is evaluated. To do this, the space power supply design process and its associated knowledge base were analyzed and characterized in a form suitable for computer emulation of a human expert. The existing expert system tools and the results achieved with them were evaluated to assess their applicability to power system design. Some new concepts for combining program architectures (modular expert systems and algorithms) with information about the domain were applied to create a 'deep' system for handling the complex design problem. NOVICE, a code to solve a simplified version of a scoping study of a wide variety of power supply types for a broad range of missions, has been developed, programmed, and tested as a concrete feasibility demonstration.

Cooper, Ralph S.↗

NASA NTRS1993

The Generic Spacecraft Analyst Assistant (gensaa): a Tool for Developing Graphical Expert Systems

During numerous contacts with a satellite each day, spacecraft analysts must closely monitor real-time data. The analysts must watch for combinations of telemetry parameter values, trends, and other indications that may signify a problem or failure. As the satellites become more complex and the number of data items increases, this task is becoming increasingly difficult for humans to perform at acceptable performance levels. At NASA GSFC, fault-isolation expert systems are in operation supporting this data monitoring task. Based on the lessons learned during these initial efforts in expert system automation, a new domain-specific expert system development tool named the Generic Spacecraft Analyst Assistant (GenSAA) is being developed to facilitate the rapid development and reuse of real-time expert systems to serve as fault-isolation assistants for spacecraft analysts. Although initially domain-specific in nature, this powerful tool will readily support the development of highly graphical expert systems for data monitoring purposes throughout the space and commercial industry.

Hughes, Peter M.↗

DOE OSTI2023

Coconut oil derived five‐component synthetic oviposition deterrent for oriental fruit fly, Bactrocera dorsalis

Abstract BACKGROUND Bactrocera dorsalis, oriental fruit fly (OFF), is one of the most destructive agricultural pests. Although bait sprays can effectively control OFF,resistance development has been a concern. We evaluated the oviposition deterrent activity of coconut free fatty acids (CFFA), a mixture of eight coconut oil‐derived fatty acids known to repel hematophagous insects and deter their feeding and oviposition, against OFF females. RESULTS In laboratory 72‐h two‐choice assays using guava‐juice infused‐agar as an oviposition substrate, CFFA deterred OFF oviposition in a dose‐dependent manner with the greatest reduction of 87% at 20 mg dose compared to the control. When the eight CFFA components were tested individually, four compounds (caprylic, capric, oleic, and linoleic acids) significantly reduced OFF oviposition (‘negative‐compounds’), two (lauric and myristic acids) had no effect (‘neutral‐compounds’), and two (palmitic and stearic acids) stimulated OFF oviposition (‘positive‐compounds’). In two‐choice tests, the ‘negative‐compounds’ blend failed to elicit the same level of oviposition reduction as CFFA at equivalent concentrations found in CFFA. Adding the two ‘neutral‐compounds’ recovered the oviposition deterrence similar to CFFA. Subsequent subtraction tests showed that four ‘negative‐compounds’ plus lauric acid was as effective as CFFA in reducing OFF oviposition in guava‐juice agar. This five‐component key‐deterrent blend also reduced OFF oviposition by 95 and 72% on papaya and tomato fruit, respectively. CONCLUSION CFFA acts as an oviposition deterrent for OFF. Given that CFFA compounds are generally regarded as safe for humans and the environment, CFFA and its bioactive components have potential use in behavioral control strategies against OFF. © 2023 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Agriculture↗

DOE OSTI2021

Structural basis for SARS-CoV-2 envelope protein recognition of human cell junction protein PALS1

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has created global health and economic emergencies. SARS-CoV-2 viruses promote their own spread and virulence by hijacking human proteins, which occurs through viral protein recognition of human targets. To understand the structural basis for SARS-CoV-2 viral-host protein recognition, here we use cryo-electron microscopy (cryo-EM) to determine a complex structure of the human cell junction protein PALS1 and SARS-CoV-2 viral envelope (E) protein. Our reported structure shows that the E protein C-terminal DLLV motif recognizes a pocket formed exclusively by hydrophobic residues from the PDZ and SH3 domains of PALS1. Our structural analysis provides an explanation for the observation that the viral E protein recruits PALS1 from lung epithelial cell junctions. In addition, our structure provides novel targets for peptide- and small-molecule inhibitors that could block the PALS1-E interactions to reduce E-mediated virulence.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Study of Protein Dynamics via Neutron Spin Echo Spectroscopy

Most human body proteins' activity and functionality are related to configurational changes of entire subdomains within the protein crystal structure. The crystal structures build the basis for any calculation that describes the structure or dynamics of a protein, most of the time with strong geometrical restrictions. However, these restrictions from the crystal structure are not present in the solution. The structure of the proteins in the solution may differ from the crystal due to rearrangements of loops or subdomains on the pico to nanosecond time scale (i.e., the internal protein dynamics time regime). This study describes how slow motions on timescales of several tens of nanoseconds can be accessed using neutron scattering. In particular, the dynamical characterization of two major human proteins, an intrinsically disordered protein that lacks a well-defined secondary structure and a classical antibody protein, is addressed by neutron spin echo spectroscopy (NSE) combined with a wide range of laboratory characterization methods. Further insights into protein domain dynamics were achieved using mathematical modeling to describe the experimental neutron data and determine the crossover between combined diffusive and internal protein motions. The extraction of the internal dynamic contribution to the intermediate scattering function obtained from NSE, including the timescale of the various movements, allows further vision into the mechanical properties of single proteins and the softness of proteins in their nearly natural environment in the crowded protein solution.

59 BASIC BIOLOGICAL SCIENCES↗