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All sourcesNASA NTRSDOE OSTI
NASA NTRS2017

Low-Melt Polyamic Acid Based Powder Coatings

The present invention is directed to a method for powder coating a metal substrate using a low-melt polyamic acid (PAA) polymer that readily imidizes to polyimides. These low-melt PAAs have been shown to be useful in resins applied as powder coatings to metal surfaces. The resin includes an end-capping material capable of providing crosslinking functionality to at least one end of the low-melt PAA polymer. The end-capping material functions dually as a polymerization chain terminator and crosslinking agent, thus producing resins that have molecular weights low enough to flow well and form good cured films applicable for use in powder coating.

Jolley, Scott T.↗
NASA NTRS2019

A Single Stage to Orbit Design for a Hybrid Mars Ascent

A Single Stage to Orbit (SSTO) hybrid propulsion system has been previously studied as an option for a conceptual Mars Ascent Vehicle (MAV). The hybrid motor uses a wax-based fuel developed specifically for this application, so it can take advantage of a single port design. The oxidizer is Mixed Oxides of Nitrogen (MON-25). Higher Nitric Oxide concentrations have been discussed in the past, however, the lower temperature capability is no longer needed. The MAV Payload Assembly (MPA), which would house the Orbiting Sample (OS) has changed substantially from previous iterations and has become more compact. Benefits of the hybrid option include its predicted low temperature behavior, high performance and ability to restart (enabling the SSTO). However, the hybrid technology remained at a relatively low Technology Readiness Level (TRL). In an attempt to increase the TRL, a technology development program has been underway for the past four years. The results of the technology development program are now being incorporated to an updated concept for a hybrid Mars Ascent Vehicle, with the eventual goal of informing a hybrid propulsion design that closes under the guidelines currently envisioned for a potential Mars Sample Return campaign. This paper focuses on the hybrid propulsion system design and the preliminary results from the first part of the FY19 technology development program (October 2018 to July 2019) and includes some results from a Preliminary Architecture Assessment (PAA) study. In the PAA, experts from all relevant subsystems (propulsion, avionics, GN&C, structures, thermal, etc.) are brought together to determine an updated vehicle design. The PAA is being run out of Marshall Space Flight Center (MSFC) in coordination with the Mars Sample Return study lead by the Jet Propulsion Laboratory (JPL). Currently, it is thought that the Mars Ascent Vehicle would be housed in a Sample Retrieval Lander (SRL), along with a fetch rover. The SRL would be responsible for several crucial functions on the MAV including heating, erection and providing the ignition signal. This paper will also outline the future testing and path forward through the rest of the fiscal year. This includes full scale testing at Whittinghill Aerospace, hypergolic additive testing at Purdue, evaluation of adding hypergolic additives to a full-scale grain. A hybrid fuel formulation has been updated with a reduced regression rate, which again was developed by Space Propulsion Group. This design will be used to determine the benefits of a hybrid versus solid propulsion system for a MAV, as they fit into the larger vision for a potential Mars Sample Return campaign.

Story, George↗
NASA NTRS2020

Constraining Remote Oxidation Capacity with ATom Observations

The global oxidation capacity, defined as the tropospheric mean concentration of the hydroxyl radical (OH), controls the lifetime of reactive trace gases in the atmosphere such as methane and carbon monoxide (CO). Models tend to underestimate the methane lifetime and CO concentrations throughout the troposphere, which is consistent with excessive OH. Approximately half the oxidation of methane and non-methane volatile organic compounds (VOCs) is thought to occur over the oceans where oxidant chemistry has received little validation due to a lack of observational constraints. We use observations from the first two deployments of the NASA ATom aircraft campaign during July–August 2016 and January–February 2017 to evaluate the oxidation capacity over the remote oceans and its representation in the GEOS-Chem chemical transport model. The model successfully simulates the magnitude and vertical profile of remote OH within the measurement uncertainties. Comparisons against the drivers of OH production (water vapor, ozone, and NOy concentrations, ozone photolysis frequencies) also show minimal bias with the exception of wintertime NOy, for which a model overestimate may indicate insufficient wet scavenging and/or missing loss on seasalt aerosol but large uncertainties remain that require further studies of NOy partitioning and removal in the troposphere. During the ATom-1 deployment, OH reactivity (OHR) below 3 km is significantly enhanced, and this is not captured by the sum of its measured components (cOHRobs) or by the model (cOHRmod). This enhancement could suggest missing reactive VOCs but cannot be explained by new estimates of ocean VOC sources and additional modeled reactivity in this region would be difficult to reconcile with the full suite of ATom measurement constraints. The model generally reproduces the magnitude and seasonality of cOHRobs but underestimates the contribution of oxygenated VOC, mainly acetaldehyde, which is severely underestimated throughout the troposphere despite its calculated lifetime of less than a day. Missing model acetaldehyde in previous studies was attributed to measurement uncertainties that have been largely resolved. Observations of peroxyacetic acid (PAA) provide new support for remote levels of acetaldehyde. The underestimate in modeled acetaldehyde and PAA is present throughout the year in both hemispheres and peaks during Northern Hemisphere summer. The addition of ocean VOC sources in the model increases annual surface cOHRmod by 10 % and improves model-measurement agreement for acetaldehyde particularly in winter but cannot resolve the model summertime bias. Doing so would require a 100 Tg yr−1 source of a long-lived unknown precursor throughout the year with significant additional emissions in the Northern Hemisphere summer. Improving the model bias for remote acetaldehyde and PAA is unlikely to fully resolve previously reported model global biases in OH and methane lifetime, suggesting that future work should examine the sources and sinks of OH over land.

Travis, Katherine R.↗
NASA NTRS1991

Plasmoids as magnetic flux ropes

A magnetic flux rope model is developed and used to determine whether the principal axis analysis (PAA) of magnetometer signatures from a single satellite pass is sufficient to obtain the magnetic topology of plasmoids. The model is also used to determine if plasmoid observations are best explained by the flux rope, closed loop, or large-amplitude wave picture. It was found that the principal axis directions is highly dependent on the satellite trajectory through the structure and, therefore, the PAA of magnetometer data from a single satellite pass is insufficient to differentiate between magnetic closed loop and flux rope models. Results also indicate that the flux rope model of plasmoid formation is well suited to unify the observations of various magnetic structures observed by ISEE 3.

Moldwin, Mark B.↗
NASA NTRS2001

Zr-Containing 4,4'-ODA/PMDA Polyimide Composites

The objective of this research is to improve the atomic oxygen resistance of Kapton(TM), a polyimide (PI) made from pyromellitic acid dianhydride (PMDA) and 4,4'-oxydianiline (ODA), while retaining or enhancing the desirable properties of the pure polymer. Toward this end, zirconium-containing complexes and polymers were used to make composites and blends. Tetra(acetylacetonato)zirconium(IV), Zr(acac)4, which is commercially available, was identified as the best zirconium-containing complex for enhancing the atomic oxygen resistance of polyimide composites of the 10 complexes screened. Films prepared from the commercially available polyamic acid (PAA) of PMDA-ODA (DuPont) have good uniformity, flexibility, and tensile strength. A 24-layer 10% (mol) Zr(acac)4/PI composite film showed significant improvement (ca. 20 fold) of atomic oxygen resistance over the pure polyimide. However, 10% (mol) Zr(acac)4 represents an upper concentration limit, above which films undergo cracking upon thermal imidization. In order to increase the Zr complex concentration in PMDA-ODA PI films, while retaining good film properties, [Zr(adsp)2-PMDA]n coordination polymer [bis(4-amino-N,N'-disalicylidene- 1,2-phenylenediamino)zirconium(IV)-pyromellitic dianhydride] and [Zr(adsp)2-PMDA-ODA-PMDA]n terpolymer were synthesized and blended with commercial PAA, respectively. Several techniques were used to characterize the films made from the polymer containing Zr(acac)4. Plasma studies of films having 2% (mol) incremental concentrations of Zr in the Kapton up to 10% (mol) show that the overall rate of erosion is reduced about 75 percent.

Illingsworth, M. L.↗
NASA NTRS2006

Reducing Propulsion Airframe Aeroacoustic Interactions With Uniquely Tailored Chevrons: 3. Jet-Flap Interaction

Propulsion airframe aeroacoustic (PAA) interactions, resulting from the integration of engine and airframe, lead to azimuthal asymmetries in the flow/acoustic field, e.g., due to the interaction between the exhaust jet flow and the pylon, the wing and its high-lift devices, such as, flaps and flaperons. In the first two parts of this series we have presented experimental results which show that isolated and installed nozzles with azimuthally varying chevrons (AVCs) can reduce noise more than conventional chevrons when integrated with a pylon and a wing with flaps at take-off conditions. In this paper, we present model-scale experimental results for the reduction of jet-flap interaction noise source due to these AVCs and document the PAA installation effects (difference in noise between installed and isolated nozzle configurations) at both approach and take-off conditions. It is found that the installation effects of both types of chevron nozzles, AVCs and conventional, are reversed at approach and take-off, in that there is more installed noise reduction at approach and less at take-off compared to that of the isolated nozzles. Moreover, certain AVCs give larger total installed noise benefits at both conditions compared to conventional chevrons. Phased microphone array results show that at approach conditions (large flap deflection, low jet speed and low ambient Mach number), chevrons gain more noise benefit from reducing jetflap interaction noise than they do from quieting the jet plume noise source which is already weak at these low jet speeds. In contrast, at take-off (small flap deflection, high jet speed and high ambient Mach number) chevrons reduce the dominant jet plume noise better than the reduction they create in jet-flap interaction noise source. In addition, fan AVCs with enhanced mixing near the pylon are found to reduce jet-flap interaction noise better than conventional chevrons at take-off.

Thomas, Russ H.↗
NASA NTRS2006

Reducing Propulsion Airframe Aeroacoustic Interactions with Uniquely Tailored Chevrons: Installed Nozzles - 2

Propulsion airframe aeroacoustic (PAA) interactions arise due to the manner in which an engine is installed on the airframe and lead to an asymmetry in the flow/acoustic environment, for example, for under-the-wing installations due to the pylon, the wing and the high-lift devices. In this work we study how we can affect these PAA interactions to reduce the overall jet-related installed noise by tailoring the chevron shapes on fan and core nozzles in a unique fashion to take advantage of this asymmetry. In part 1 of this trio of papers we introduced the concept of azimuthally varying chevrons (AVC) and showed how some types of AVCs can be more beneficial than the conventional chevrons when tested on "isolated" scaled nozzles inclusive of the pylon effect. In this paper, we continue to study the effect of installing these AVC nozzles under a typical scaled modern wing with high-lift devices placed in a free jet. The noise benefits of these installed nozzles, as well as their installation effects are systematically studied for several fan/core AVC combinations at typical take-off conditions with high bypass ratio. We show, for example, that the top-enhanced mixing T-fan AVC nozzle (with enhanced mixing near the pylon and less mixing away from it) when combined with conventional chevrons on the core nozzle is quieter than conventional chevrons on both nozzles, and hardly produces any high-frequency lift, just as in the isolated case; however, its installed nozzle benefit is less than its isolated nozzle benefit. This suppression of take-off noise benefit under installed conditions, compared to its isolated nozzle benefit, is seen for all other chevron nozzles. We show how these relative noise benefits are related to the relative installation effects of AVCs and baseline nozzles.

Mengle, Vinod G.↗
NASA NTRS2014

Vertical Carbon Nanotube Device in Nanoporous Templates

A modified porous anodic alumina template (PAA) containing a thin CNT catalyst layer directly embedded into the pore walls. CNT synthesis using the template selectively catalyzes SWNTs and DWNTs from the embedded catalyst layer to the top PAA surface, creating a vertical CNT channel within the pores. Subsequent processing allows for easy contact metallization and adaptable functionalization of the CNTs and template for a myriad of applications.

Maschmann, Matthew Ralph↗
NASA NTRS2018

Aircraft System Noise Assessment of the NASA D8 Subsonic Transport Concept

Avehicle-level noise assessment has been performed for theNASAD8 concept aircraft (ND8) in the NASA Advanced Air Transport Technology Project portfolio. The NASA research-level Aircraft NOise Prediction Program (ANOPP-Research) was used to predict the noise from each source component on the ND8 to build up a noise estimate for the full aircraft. The propulsion airframe aeroacoustic (PAA) effects of the ND8, namely boundary layer ingestion (BLI) with its influence on fan noise, and the noise shielding, reflection, and diffraction mechanisms of the unconventional airframe, were empirically modeled using experimental data. Noise reduction technologies appropriate to the 2025-2035 time frame were included in this study. Including all technologies and PAA effects, the ND8 is predicted to have a cumulative margin to the Stage 4 certification metric of only 7.4 EPNdB. Boundary layer ingestion is predicted to have a detrimental impact on cumulative noise levels on the order of 15 EPNdB. Fan noise is seen to be the primary noise source at all three certification points, even if the BLI noise impact could be entirely suppressed. The impact of engine noise shielding by the airframe is limited by a lack of aft shielding and the presence of horizontal tail reflections in the aft direction. The physical constraint on engine size by the pi-tail is seen as a potential barrier to engine noise reduction through the corresponding limitation on fan bypass ratio. Mildly reduced climb performance (compared to similar reference aircraft) does not provide any benefit through increased noise propagation distance. If the boundary layer ingestion noise penalty could be suppressed such that BLI would have no effect on noise, the cumulative margin to Stage 4 would increase to 22.4 EPNdB, still below the NASA Mid Term goal of 32-42 EPNdB.

Clark, Ian A.↗
NASA NTRS2019

Mars-GRAM Support for the Mars Ascent Vehicle

Mars sample return is a bold concept, which entails gathering a varied, scientifically-relevant collection of Martian rock core samples and bringing them to Earth for analysis. To support this endeavor, the Marshall Space Flight Center (MSFC) is developing the Mars Ascent Vehicle (MAV), which is responsible for getting the collected samples off the planet. The MAV Preliminary Architecture Assessment (PAA) study is designing two vehicle architectures based on different propulsion configurations: a two-stage solid-solid concept, and a hybrid concept. Given different thrust profiles for the two configurations, each concept uses a unique trajectory to reach the same orbit. In support of the PAA, The MSFC Natural Environments Branch (EV44) was asked to produce tables of atmospheric parameters along each of the two trajectories. The Mars Global Reference Atmospheric Model (Mars-GRAM) is an EV44 tool that is ideally suited for this analysis. Mars-GRAM will continue supporting MAV development in future design cycles.

Burns, K. L.↗
NASA NTRS2020

Mars Ascent Vehicle Hybrid Propulsion Configuration

As part of a Mars Sample Return (MSR) campaign, two Mars Ascent Vehicle (MAV) configurations have been designed in parallel. Each ascent vehicle configuration has a different propulsion system which ultimately leads to two unique vehicle designs. As part of a Preliminary Architecture Assessment (PAA), these vehicle designs were developed to the same level of maturity in order to inform the selection of one of the vehicles as the point of departure design for the campaign. The selection will be made in November 2019. The initial MSR architecture called for a hybrid-based propulsion MAV. This type of propulsion system calls for a solid wax motor that would utilize liquid MON-25 as an oxidizer. Hybrid rocket propulsion allows for more flexibility than traditional solid or liquid propulsion options, and typically benefits from the advantages of both. A hybrid motor can be throttled and shut down easily, and avoids significant risk in manufacturing and handling. On a theoretical level, hybrid motors perform at a higher specific impulse (Isp) than solid motors. The primary disadvantage of hybrid motors comes from additional complexity and significantly less flight heritage and low Technology Readiness Level (TRL). This paper describes the design of the hybrid propulsion configuration. An additional paper will be published describing the design of the solid propulsion configuration1. The hybrid propulsion configuration MAV was developed in 2019 by NASA Marshall Space Flight Center (MSFC) in association with NASA Jet Propulsion Laboratory (JPL). It features a Single Stage to Orbit (SSTO) design with an SP7A solid wax fuel and MON-25 liquid oxidizer. The liquid portion of the vehicle allows for a Liquid Injection Thrust Vector Controller (LITVC) as well as hypergolic propellant additives for ignition. The vehicle was designed to deliver approximately 0.31kg of Martian geological samples to a circular orbit at Mars of 343km at a 25o inclination. Although hybrid propulsion in general has been used on launch vehicles in the past, the integrated vehicle subsystems that operate in conjunction with these propulsion elements do not typically operate in a Martian environment, which in this application can get as cold as -40oC. The PAA advanced the maturity of these subsystems by performing detailed design and analysis on the vehicle with respect to structures and mechanisms, Guidance/Navigation/Control (GNC) systems, avionics, Reaction Control System (RCS), LITVC, thermal environments, and advanced Computational Fluid Dynamics (CFD). This paper will summarize the results of these studies.

Yaghoubi, Darius↗
NASA NTRS2020

Mars Ascent Vehicle Solid Propulsion Configuration

As part of a Mars Sample Return (MSR) campaign, two Mars Ascent Vehicle (MAV) configurations have been designed in parallel. Each ascent vehicle configuration has a different propulsion system, which ultimately leads to two unique vehicle designs. As part of a Preliminary Architecture Assessment (PAA), these vehicle designs were developed to the same level of maturity in order to inform the selection of one of the vehicles as the point of departure design for the campaign. The selection will be made in November 2019. Although the initial MSR architecture called for a hybrid-based propulsion MAV featuring solid wax fuel with liquid oxidizer, a configuration using more traditional solid propulsion was developed as an additional risk mitigation option. Though lacking in the single stage to orbit (SSTO) and throttle flexibility of a hybrid configuration, a solid configuration vehicle allows a simpler design with significantly longer flight heritage and higher Technology Readiness Level (TRL). This paper describes the design of the solid propulsion configuration. An additional paper will be published describing the design of the hybrid propulsion configuration. The solid propulsion configuration MAV was developed in 2019 by NASA Marshall Space Flight Center (MSFC) in association with NASA Jet Propulsion Laboratory (JPL). It features two stages with a modified STAR-17 motor for the second stage and a traditional electromechanical actuator Thrust Vector Controller (TVC). The vehicle was designed to deliver approximately 0.47kg of Martian geological samples to a circular orbit at Mars of 343km at a 25° inclination. Although solid motor designs in general are at a relatively high TRL, the integrated vehicle subsystems that operate in conjunction with these propulsion elements do not typically operate in a Martian environment, which in this application can get as cold as -40°C. The PAA advanced the maturity of these subsystems by performing detailed design and analysis on the vehicle with respect to structures and mechanisms, Guidance/Navigation/Control (GNC) systems, avionics, Reaction Control System (RCS), TVC, thermal environments, and advanced Computational Fluid Dynamics (CFD). This paper will summarize the results of these studies.

Yaghoubi, Darius↗
NASA NTRS2020

Mars Ascent Vehicle Solid Propulsion Configuration

As part of a Mars Sample Return (MSR) campaign, two Mars Ascent Vehicle (MAV) configurations have been designed in parallel. Each ascent vehicle configuration has a different propulsion system, which ultimately leads to two unique vehicle designs. As part of a Preliminary Architecture Assessment (PAA), these vehicle designs were developed to the same level of maturity in order to inform the selection of one of the vehicles as the point of departure design for the campaign. The selection will be made in November 2019. Although the initial MSR architecture called for a hybrid-based propulsion MAV featuring solid wax fuel with liquid oxidizer, a configuration using more traditional solid propulsion was developed as an additional risk mitigation option. Though lacking in the single stage to orbit (SSTO) and throttle flexibility of a hybrid configuration, a solid configuration vehicle allows a simpler design with significantly longer flight heritage and higher Technology Readiness Level (TRL). This paper describes the design of the solid propulsion configuration. An additional paper will be published describing the design of the hybrid propulsion configuration. The solid propulsion configuration MAV was developed in 2019 by NASA Marshall Space Flight Center (MSFC) in association with NASA Jet Propulsion Laboratory (JPL). It features two stages with a modified STAR-17 motor for the second stage and a traditional electromechanical actuator Thrust Vector Controller (TVC). The vehicle was designed to deliver approximately 0.47kg of Martian geological samples to a circular orbit at Mars of 343km at a 25° inclination. Although solid motor designs in general are at a relatively high TRL, the integrated vehicle subsystems that operate in conjunction with these propulsion elements do not typically operate in a Martian environment, which in this application can get as cold as -40°C. The PAA advanced the maturity of these subsystems by performing detailed design and analysis on the vehicle with respect to structures and mechanisms, Guidance/Navigation/Control (GNC) systems, avionics, Reaction Control System (RCS), TVC, thermal environments, and advanced Computational Fluid Dynamics (CFD). This paper will summarize the results of these studies.

Yaghoubi, Darius↗
NASA NTRS1981

Demonstration Advanced Avionics System (DAAS), Phase 1

Demonstration advanced anionics system (DAAS) function description, hardware description, operational evaluation, and failure mode and effects analysis (FMEA) are provided. Projected advanced avionics system (PAAS) description, reliability analysis, cost analysis, maintainability analysis, and modularity analysis are discussed.

Bailey, A. J.↗
NASA NTRS1988

Chemistry of the older supracrustals of Archaean age around Sargur

In the Archaeans of the Karnataka craton two stratigraphically distinct volcano-sedimentary sequences occur, namely the older supracrustals of the Sargur type and the younger Dharwar greenstones. The dividing line between these is the 3 by old component of the Peninsular gneiss. The trace and rare earth element chemistry of the Sargur metasediments show, in general, marked similarity to the Archaean sediments. The significant departures are in the nickel and chromium abundances. The REE data of the Sargur pelites of the Terakanambi region represented by Silli-gt-bio-feldspar schists and paragneisses show LREE enrichment and flat to depleted HREE pattern. Banded iron formations have very low REE abundance. They show slightly enriched LREE and flat to depleted HREE pattern. REE abundance in the Mn-horizons is comparable to that of the Archaean sediments. Mn-horizons show enriched LREE and flat HREE with anamolous Eu. REE patterns of these bands is well evolved and has similarities with PAAS.

Janardhan, A. S.↗
NASA NTRS1991

Knowledge-based decision support for Space Station assembly sequence planning

A complete Personal Analysis Assistant (PAA) for Space Station Freedom (SSF) assembly sequence planning consists of three software components: the system infrastructure, intra-flight value added, and inter-flight value added. The system infrastructure is the substrate on which software elements providing inter-flight and intra-flight value-added functionality are built. It provides the capability for building representations of assembly sequence plans and specification of constraints and analysis options. Intra-flight value-added provides functionality that will, given the manifest for each flight, define cargo elements, place them in the National Space Transportation System (NSTS) cargo bay, compute performance measure values, and identify violated constraints. Inter-flight value-added provides functionality that will, given major milestone dates and capability requirements, determine the number and dates of required flights and develop a manifest for each flight. The current project is Phase 1 of a projected two phase program and delivers the system infrastructure. Intra- and inter-flight value-added were to be developed in Phase 2, which has not been funded. Based on experience derived from hundreds of projects conducted over the past seven years, ISX developed an Intelligent Systems Engineering (ISE) methodology that combines the methods of systems engineering and knowledge engineering to meet the special systems development requirements posed by intelligent systems, systems that blend artificial intelligence and other advanced technologies with more conventional computing technologies. The ISE methodology defines a phased program process that begins with an application assessment designed to provide a preliminary determination of the relative technical risks and payoffs associated with a potential application, and then moves through requirements analysis, system design, and development.

Source record↗
NASA NTRS1993

Surface analyses of composites exposed to the space environment on LDEF

A series of surface analyses on carbon fiber/poly(arylacetylene) (PAA) matrix composites that were exposed to the space environment on the Long Duration Exposure Facility (LDEF) satellite were conducted. These composite panels were arranged in pairs on both the leading edge and trailing edge of LDEF. None of the composites were catastrophically damaged by nearly six years of exposure to the space environment. Composites on the leading edge exhibited from 25 to 125 microns of surface erosion, but trailing edge panels exhibited no physical appearance changes due to exposure. Scanning electron microscopy (SEM) was used to show that the erosion morphology on the leading edge samples was dominated by crevasses parallel to the fibers with triangular cross sections 10 to 100 microns in depth. The edges of the crevasses were well defined and penetrated through both matrix and fiber. The data suggest that the carbon fibers are playing an important role in crevasse initiation and/or enlargement, and in the overall erosion rate of the composite. X-ray photoelectron spectroscopy (XPS) and energy dispersive x-ray spectroscopy (EDS) results showed contamination from in-flight sources of silicone.

Mallon, Joseph J.↗
NASA NTRS1999

Fluorescence Studies of Protein Crystal Nucleation

Fluorescence can be used to study protein crystal nucleation through methods such as anisotropy, quenching, and resonance energy transfer (FRET), to follow pH and ionic strength changes, and follow events occurring at the growth interface. We have postulated, based upon a range of experimental evidence that the growth unit of tetragonal hen egg white lysozyme is an octamer. Several fluorescent derivatives of chicken egg white lysozyme have been prepared. The fluorescent probes lucifer yellow (LY), cascade blue, and 5-((2-aminoethyl)aminonapthalene-1-sulfonic acid (EDANS), have been covalently attached to ASP 101. All crystallize in the characteristic tetragonal form, indicating that the bound probes are likely laying within the active site cleft. Crystals of the LY and EDANS derivatives have been found to diffract to at least 1.7 A. A second group of derivatives is to the N-terminal amine group, and these do not crystallize as this site is part of the contact region between the adjacent 43 helix chains. However derivatives at these sites would not interfere with formation of the 43 helices in solution. Preliminary FRET studies have been carried out using N-terminal bound pyrene acetic acid (Ex 340 nm, Em 376 nm) lysozyme as a donor and LY (Ex -425 nm, Em 525 nm) labeled lysozyme as an acceptor. FRET data have been obtained at pH 4.6, 0.1 M NaAc buffer, at 5 and 7% NaCl, 4 C. The corresponding Csat values are 0.471 and 0.362 mg/ml (approximately 3.3 and approximately 2.5 x 10(exp -5) M respectively). The data at both salt concentrations show a consistent trend of decreasing fluorescence intensity of the donor species (PAA) with increasing total protein concentration. This decrease is more pronounced at 7% NaCl, consistent with the expected increased intermolecular interactions at higher salt concentrations reflected in the lower solubility. The calculated average distance between any two protein molecules at 5 x 10(exp -6) M is approximately 70nm, well beyond the range where any FRET can be expected. Results from these and ongoing studies will be presented.

Pusey, Marc L.↗