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At least 541 records · Page 30
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

Signal sequences target enzymes and structural proteins to bacterial microcompartments and are critical for microcompartment formation

ABSTRACT Spatial organization of pathway enzymes has emerged as a promising tool to address several challenges in metabolic engineering, such as flux imbalances and off-target product formation. Bacterial microcompartments (MCPs) are a spatial organization strategy used natively by many bacteria to encapsulate metabolic pathways that produce toxic, volatile intermediates. Several recent studies have focused on engineering MCPs to encapsulate heterologous pathways of interest, but how this engineering affects MCP assembly and function is poorly understood. In this study, we investigated the role of signal sequences, short domains that target proteins to the MCP core, in the assembly of 1,2-propanediol utilization (Pdu) MCPs. We characterized two novel Pdu signal sequences on the structural proteins PduM and PduB, which constitute the first report of metabolosome signal sequences on structural proteins rather than enzymes. We then explored the role of enzymatic and structural Pdu signal sequences on MCP assembly by deleting their encoding sequences from the genome alone and in combination. Deleting enzymatic signal sequences decreased the MCP formation, but this defect could be recovered in some cases by overexpressing genes encoding the knocked-out signal sequence fused to a heterologous protein. By contrast, deleting structural signal sequences caused similar defects to knocking out the genes encoding the full-length PduM and PduB proteins. Our results contribute to a growing understanding of how MCPs form and function in bacteria and provide strategies to mitigate assembly disruption when encapsulating heterologous pathways in MCPs. IMPORTANCE Spatially organizing biosynthetic pathway enzymes is a promising strategy to increase pathway throughput and yield. Bacterial microcompartments (MCPs) are proteinaceous organelles that many bacteria natively use as a spatial organization strategy to encapsulate niche metabolic pathways, providing significant metabolic benefits. Encapsulating heterologous pathways of interest in MCPs could confer these benefits to industrially relevant pathways. Here, we investigate the role of signal sequences, short domains that target proteins for encapsulation in MCPs, in the assembly of 1,2-propanediol utilization (Pdu) MCPs. We characterize two novel signal sequences on structural proteins, constituting the first Pdu signal sequences found on structural proteins rather than enzymes, and perform knockout studies to compare the impacts of enzymatic and structural signal sequences on MCP assembly. Our results demonstrate that enzymatic and structural signal sequences play critical but distinct roles in Pdu MCP assembly and provide design rules for engineering MCPs while minimizing disruption to MCP assembly.

Johnson, Elizabeth R. (ORCID:0000000179236881)↗
DOE OSTI2025

Structural and functional analyses of SARS-CoV-2 Nsp3 and its specific interactions with the 5’ UTR of the viral genome

ABSTRACT Non-structural protein 3 (Nsp3) is the largest open reading frame encoded in the SARS-CoV-2 genome, essential for the formation of double-membrane vesicles (DMV) wherein viral RNA replication occurs. We conducted an extensive structure-function analysis of Nsp3 and determined the crystal structures of the ubiquitin-like 1 (Ubl1), nucleic acid binding (NAB), β-coronavirus-specific marker (βSM) domains, and a sub-region of the Y domain of this protein. We show that the Ubl1, ADP-ribose phosphatase (ADRP), human SARS Unique (HSUD), NAB, and Y domains of Nsp3 bind the 5’ UTR of the viral genome and that the Ubl1 and Y domains possess affinity for recognition of this region, suggesting high specificity. The Ubl1-Nucleocapsid (N) protein complex binds the 5’ UTR with greater affinity than the individual proteins alone. Our results suggest that multiple domains of Nsp3, particularly Ubl1 and Y, shepherd the 5’ UTR of the viral genome during translocation through the DMV membrane, priming the Ubl1 domain to load the genome onto N protein. IMPORTANCE The largest protein encoded by the SARS-CoV-2 genome is Nsp3. In infected cells, this multi-domain protein forms a pore structure in the virus-induced double-membrane vesicles (DMV). We have incomplete data on Nsp3 molecular structure, and here, we describe crystal structures for multiple domains of Nsp3. It is thought that newly replicated viral RNA transits through the DMV pore; however, we possess incomplete data on which regions of Nsp3 actually interact with RNA. Here, we present data showing that five domains of Nsp3 interact with the 5’ UTR of the SARS-CoV-2 RNA, including the Y domain for which no function has ever been discovered. These data suggest that the pore structure plays an active role in recognizing the terminal end of the genome, transiting and loading the viral RNA onto the cytoplasmic nucleocapsid protein. These data help expand our knowledge of Nsp3 structure and function and the SARS-CoV-2 replication cycle.

Microbiology↗
DOE OSTI2025

Confocal Raman Microscopy for Measuring In Situ Temperature-Dependent Structural Changes in Poly(Ethylene Oxide) Thin Films

Crystallization from the melt is a critical process governing the properties of semi-crystalline polymeric materials. While structural analyses of melting and crystallization transitions in bulk polymers have been widely reported, in contrast, those in thin polymer films on solid supports have been underexplored. Herein, in situ Raman microscopy and self-modeling curve resolution (SMCR) analysis are applied to investigate the temperature-dependent structural changes in poly(ethylene oxide) (PEO) films during melting and crystallization phase transitions. By resolving complex overlapping sets of spectra, SMCR analysis reveals that the thermal transitions of 50 µm thick PEO films comprise two structural phases: an ordered crystalline phase and a disordered amorphous phase. The ordered structure of the crystalline PEO film entirely disappears as the polymer is heated; conversely, the disordered structure of the amorphous PEO film reverts to the ordered structure as the polymer is cooled. Broadening of the Raman bands was observed in PEO films above the melting temperature (67 °C), while sharpening of bands was observed below the crystallization temperature (45 °C). The temperatures at which these spectral changes occurred were in good agreement with differential scanning calorimetry (DSC) measurements, especially during the melting transition. The results illustrate that in situ Raman microscopy coupled with SMCR analysis is a powerful approach for unraveling complex structural changes in thin polymer films during melting and crystallization processes. Furthermore, we show that confocal Raman microscopy opens opportunities to apply the methodology to interrogate the structural features of PEO or other surface-supported polymer films as thin as 2 µm, a thickness regime beyond the reach of conventional thermal analysis techniques.

Koh, Miharu [Department of Chemistry, University o↗
DOE OSTI2026

Doping Metallic Grain Boundaries to Control Atomic Structure and Damage Tolerance

Grain boundaries often act as sites for crack and void nucleation during plastic deformation of metallic materials. While it is known that grain boundary character and structure can greatly influence this damage nucleation process, the current level of control over such details is limited. The objective of this project was to obtain a fundamental understanding of how metallic grain boundary structure can be controlled through intelligent doping, with the idea of inducing planned amorphous grain boundary phases or complexions. The effect of amorphous complexion structure on dislocation accommodation mechanisms was studied, to improve the field’s understanding of damage nucleation at a promising type of interface. Different microstructural descriptors were studied, as grain boundaries can have large variations and complexity of local structure, and influence the mechanical damage resistance of these features was tested. Nanocrystalline systems that contain amorphous grain boundary complexions were prioritized, as these have more extreme variations in interfacial structure and damage tolerance, with an emphasis on isolating the importance of complexion population (type, thickness, etc.), network topology, local structure, and local chemistry. This research used a combination of computational, experimental, and characterization techniques to isolate and understand the importance of nanoscale grain boundary structure and interfacial chemistry. Amorphous grain boundary complexions were found to clearly increase a material’s resistance to mechanical damage, with the local distribution of structural short-range order within the complexions found to be an important descriptor for damage and the proposed focus of future work in this area.

36 MATERIALS SCIENCE↗
NASA NTRS1971

Structural interaction with control systems

A monograph which assesses the state of the art of space vehicle design and development is presented. The monograph presents criteria and recommended practices for determining the structural data and a mathematical structural model of the vehicle needed for accurate prediction of structure and control-system interaction; for design to minimize undesirable interactions between the structure and the control system; and for determining techniques to achieve the maximum desirable interactions and associated structural design benefits. All space vehicles are treated, including launch vehicles, spacecraft, and entry vehicles. Important structural characteristics which affect the structural model used for structural and control-system interaction analysis are given.

Noll, R. B.↗
NASA NTRS1977

Analysis of simple 2-D and 3-D metal structures subjected to fragment impact

Theoretical methods were developed for predicting the large-deflection elastic-plastic transient structural responses of metal containment or deflector (C/D) structures to cope with rotor burst fragment impact attack. For two-dimensional C/D structures both, finite element and finite difference analysis methods were employed to analyze structural response produced by either prescribed transient loads or fragment impact. For the latter category, two time-wise step-by-step analysis procedures were devised to predict the structural responses resulting from a succession of fragment impacts: the collision force method (CFM) which utilizes an approximate prediction of the force applied to the attacked structure during fragment impact, and the collision imparted velocity method (CIVM) in which the impact-induced velocity increment acquired by a region of the impacted structure near the impact point is computed. The merits and limitations of these approaches are discussed. For the analysis of 3-d responses of C/D structures, only the CIVM approach was investigated.

Witmer, E. A.↗
NASA NTRS1977

Study of metallic structural design concepts for an arrow wing supersonic cruise configuration

A structural design study was made, to assess the relative merits of various metallic structural concepts and materials for an advanced supersonic aircraft cruising at Mach 2.7. Preliminary studies were made to ensure compliance of the configuration with general design criteria, integrate the propulsion system with the airframe, select structural concepts and materials, and define an efficient structural arrangement. An advanced computerized structural design system was used, in conjunction with a relatively large, complex finite element model, for detailed analysis and sizing of structural members to satisfy strength and flutter criteria. A baseline aircraft design was developed for assessment of current technology. Criteria, analysis methods, and results are presented. The effect on design methods of using the computerized structural design system was appraised, and recommendations are presented concerning further development of design tools, development of materials and structural concepts, and research on basic technology.

Turner, M. J.↗
NASA NTRS1979

Weight optimization of ultra large space structures

The paper describes the optimization of a solar power satellite structure for minimum mass and system cost. The solar power satellite is an ultra large low frequency and lightly damped space structure; derivation of its structural design requirements required accommodation of gravity gradient torques which impose primary loads, life up to 100 years in the rigorous geosynchronous orbit radiation environment, and prevention of continuous wave motion in a solar array blanket suspended from a huge, lightly damped structure subject to periodic excitations. The satellite structural design required a parametric study of structural configurations and consideration of the fabrication and assembly techniques, which resulted in a final structure which met all requirements at a structural mass fraction of 10%.

Reinert, R. P.↗
NASA NTRS1983

Structures and subsystems

New and improved materials combined with efficient structural design concepts have made an essential contribution towards the shaping of the present transportation system (STS), and now, for the replacement of the STS in the year 2005, new materials and novel designs are being studied to identify the technologies which should be developed for a low-cost future space transportation system (FSTS). Three basic structural arrangements were considered for the FSTS orbiter. They include a nonintegral tank arrangement, an integral-tank arrangement, and a hybrid of the first two. Three representative arrangements regarding wall constructions are considered. Each employs a blade-stiffened aluminum tank with reinforced closed-cell-foam cryogenic insulation. Attention is given to an aluminum-alloy structure, a graphite-epoxy structure, a graphite-polyimide structure, a carbon-carbon surface panel structure, a graphite-composite fuselage structure, serviceability and all-weather considerations, and structural concept ratings.

Taylor, A. H.↗
NASA NTRS1985

Computational structural mechanics: A new activity at the NASA Langley Research Center

Complex structures considered for the late 1980's and early 1990's include composite primary aircraft structures and the space station. These structures are much more difficult to analyze than today's structures and necessitate a major upgrade in computerized structural analysis technology. A major research activity in computational structural mechanics (CSM) was initiated. The objective of the CSM activity is develop advanced structural analysis technology that will exploit modern and emerging computers such as computers with vector and/or parallel processing capabilities. The three main research activities underway in CSM include: (1) structural analysis methods development; (2) a software testbed for evaluating the methods; and (3) numerical techniques for parallel processing computers. The motivation and objectives of the CSM activity are presented and CSM activity is described. The current CSM research thrusts, and near and long term CSM research thrusts are outlined.

Knight, N. F., Jr.↗
NASA NTRS1986

Propeller-induced structure-borne noise - Laboratory-based test apparatus

A potentially important source of structure-borne interior noise transmission in advanced turboprop aircraft is the impingement of the propeller wake/vortex on downstream aerodynamic surfaces. The expected levels of propeller wake/vortex-induced structure-borne noise transmission are not known nor can they be determined with present-day technology. A test apparatus has been designed, built and calibrated for the purposes of studying propeller-induced, structure-borne noise transmission in prototypical aircraft structures. The principal approach to the test apparatus design was to provide a physical means of separating the airborne and structure-borne noise components so that the structure-borne noise transmission response could be studied directly without airborne noise contamination. This was accomplished by housing the receiving fuselage structure in an acoustic shield fitted with a wing-to-fuselage acoustic seal. Initial evaluation of the wing-to-fuselage acoustic seal indicates adequate airborne noise isolation to allow direct study of structure-borne noise transmission.

Unruh, J. F.↗
NASA NTRS1986

Design considerations for joints in deployable space truss structures

All of the structures considered for the Control of Flexible Structures (COFS) flight experiments are deployable truss structures and their response will be dominated by the structural response of the joints. To prepare for these experiments some fundamental research work is being conducted in the Structures and Dynamics Division at LaRC which will provide insight into structurally efficient and predictable deployable truss joints. This work involves generic studies of the static and dynamic response of joints as well as the development of analytical models which can be used to predict the response of a large multijointed truss. In addition to the generic joint studies, the research effort encompasses the design and fabrication of a 20-meter long deployable truss beam for laboratory evaluation of its structural characteristics and correlation with developed prediction methods. The experimental results have indicated the importance of attention to detail in the design and fabrication of joints for deployable truss structures. The dimensional relations and material considerations for efficient pin-clevis joints have been outlined. Results of tests on the near-center latch are discussed.

Rhodes, Marvin D.↗
NASA NTRS1987

Optimization procedure to control the coupling of vibration modes in flexible space structures

As spacecraft structural concepts increase in size and flexibility, the vibration frequencies become more closely-spaced. The identification and control of such closely-spaced frequencies present a significant challenge. To validate system identification and control methods prior to actual flight, simpler space structures will be flown. To challenge the above technologies, it will be necessary to design these structures with closely-spaced or coupled vibration modes. Thus, there exists a need to develop a systematic method to design a structure which has closely-spaced vibration frequencies. This paper describes an optimization procedure which is used to design a large flexible structure to have closely-spaced vibration frequencies. The procedure uses a general-purpose finite element analysis program for the vibration and sensitivity analyses and a general-purpose optimization program. Results are presented from two studies. The first study uses a detailed model of a large flexible structure to design a structure with one pair of closely-spaced frequencies. The second study uses a simple equivalent beam model of a large flexible structure to obtain a design with two pairs of closely-spaced frequencies.

Walsh, Joanne L.↗
NASA NTRS1987

Optimization procedure to control the coupling of vibration modes in flexible space structures

As spacecraft structural concepts increase in size and flexibility, the vibration frequencies become more closely-spaced. The identification and control of such closely-spaced frequencies present a significant challenge. To validate system identification and control methods prior to actual flight, simpler space structures will be flown. To challenge the above technologies it will be necessary to design these structures with closely-spaced or coupled vibration modes. Thus there exists a need to develop a systematic method to design a structure which has closely-spaced vibration frequencies. This paper describes an optimization procedure which is used to design a large flexible structure to have closely-spaced vibration frequencies. The procedure uses a general-purpose finite-element analysis program for the vibration and sensitivity analyses and a general-purpose optimization program. Results are presented from two studies. The first study uses a detailed model of a large flexible structure to design a structure with one pair of closely-spaced frequencies. The second study uses a simple equivalent beam model of a large flexible structure to obtain a design with two pairs of closely-spaced frequencies.

Walsh, Joanne L.↗
NASA NTRS1988

Large-scale wind turbine structures

The purpose of this presentation is to show how structural technology was applied in the design of modern wind turbines, which were recently brought to an advanced stage of development as sources of renewable power. Wind turbine structures present many difficult problems because they are relatively slender and flexible; subject to vibration and aeroelastic instabilities; acted upon by loads which are often nondeterministic; operated continuously with little maintenance in all weather; and dominated by life-cycle cost considerations. Progress in horizontal-axis wind turbines (HAWT) development was paced by progress in the understanding of structural loads, modeling of structural dynamic response, and designing of innovative structural response. During the past 15 years a series of large HAWTs was developed. This has culminated in the recent completion of the world's largest operating wind turbine, the 3.2 MW Mod-5B power plane installed on the island of Oahu, Hawaii. Some of the applications of structures technology to wind turbine will be illustrated by referring to the Mod-5B design. First, a video overview will be presented to provide familiarization with the Mod-5B project and the important components of the wind turbine system. Next, the structural requirements for large-scale wind turbines will be discussed, emphasizing the difficult fatigue-life requirements. Finally, the procedures used to design the structure will be presented, including the use of the fracture mechanics approach for determining allowable fatigue stresses.

Spera, David A.↗
NASA NTRS1986

Review of convectively cooled structures for hypersonic flight

Resurgent interest in development of Aerospace Plane and Orient Express type vehicles promises to stretch structural technology for hypersonic flight vehicles to the uppermost limits. Significant portions of the structure may require active cooling of some type to survive hostile environments. Despite a lack of recent research activity for cooled structures, a significant body of unclassified knowledge exists concerning such structures. Contractual and in-house research conducted mainly by NASA's Langley Research Center during the 60's and 70's on vehicles very similar to the proposed Orient Express has provided a substantial data base for convectively cooled hypersonic flight structures. Specifically, results are presented for regeneratively cooled structural concepts which have a relatively high heat flux capability and use the hydrogen fuel directly as a coolant; and for structural concepts which use a secondary coolant loop to absorb incident heating and then transfer the absorbed heat to the liquid hydrogen fuel as it flows to the engines. Results are presented to indicate application regions in terms of heat flux capability for various concepts and benefits for each concept. Experience gained and costs are discussed in terms of heat flux capability for various concepts and benefits for each concept. Additionally, experience gained and cost involved with design, fabrication, and testing of full-scale convectively cooled structures are discussed.

Shore, Charles P.↗
NASA NTRS1988

Structural tailoring and feedback control synthesis - An interdisciplinary approach

Structural tailoring provides an attractive method to optimize the performance of actively controlled space structures. However, the simultaneous optimization of control gains and structural properties often becomes prohibitively expensive for large systems and physical insight is often lost in the resulting control law. This paper presents a method for optimization of the closed loop structural system using only structural tailoring. Optimal Linear Quadratic Regulator (LQR) control theory is used with weighting matrices chosen based on physical considerations. The LQR control law depends only on two scalar gains and the structural properties. Hence, the closed loop-performance can be expressed in terms of the structural parameters. Results are given for a beam and a truss-beam to show the simplicity of the method and the importance of structural tailoring to increase dynamic performance and to reduce the control effort.

Belvin, W. Keith↗
NASA NTRS1988

Experimental studies of active members in control of large space structures

Intelligent structures for precision spacecraft applications are structural systems incorporating sensors, actuators, and built-in electronic logic that facilitate self-monitoring of structure responses to disturbances. Attention is presently given to adaptive structures, in which the electronic logic need not be integral to the structure. Active members are used to replace selected passive members of truss-type structures; the sensors employed allow measurement of the elastic strain and deformation experienced by the structure. Two struss-type testbed structures are described.

Fanson, J. L.↗