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At least 451 records · Page 25

Software Searches for Better Spacecraft-Navigation Models

ADAPT is a computer program that searches for better mathematical models for spacecraft navigation. The task of tuning trajectory-determination models for interplanetary navigation is complex, requiring an intensive search of multiple dynamical and nondynamical models that yield trajectory solutions with minimal errors. By automating the search, ADAPT eases the task of human analysts and enables them to consider wider ranges of potential solutions. ADAPT uses genetic algorithms to search a range of relevant parameters in a user-selected design space to arrive at values for those parameters that best fit the measured spacecraft-tracking data. The user s guide for ADAPT reviews the theoretical basis of the program and presents two example applications. One example is that of selecting a solar-radiation model for the Mars Pathfinder (MPF) mission using MPF tracking data and an extended Kalman filter from prior spacecraft-navigation software. The second example is of the use of tracking data from the Stardust spacecraft mission combined with a pseudo-epoch-state batch filter and an empirical small-forces model to find improved impulse models for use during Stardust attitude adjustments.

Ely, Todd↗

A Novel Multi-Spacecraft Interplanetary Global Trajectory Optimization Transcription

As the frontier of space exploration continues to advance, so does the design complexity of future interplanetary missions. One avenue of this increasing complexity includes a class of designs known as "Distributed Spacecraft Missions"; missions where multiple spacecraft coordinate to perform shared objectives. Current approaches for the global trajectory optimization of these Multi-Vehicle Missions (MVMs) are prone to shortcomings including laborious iterative design, considerable human-in-the-loop effort, treatment of the multi-vehicle problem as multiple separate trajectory optimization subproblems (resulting in suboptimal solutions where the whole is less than the sum of its parts), and poor handling of coordination objectives and constraints. There are only a handful of software platforms in existence capable of fully-automated, rapid, interplanetary mission and systems global optimization including the Parallel Global Multiobjective Optimizer (PaGMO), the Gravity Assisted Low-thrust Local Optimization Program (GALLOP), and the Evolutionary Mission Trajectory Generator (EMTG). However, none of these tools is capable of performing such tasks for MVM designs. The work outlined in this paper lays the groundwork for a technique to begin addressing these shortcomings. We present a fully-automated technique which frames interplanetary MVMs as Multi-Objective, Multi-Agent Hybrid Optimal Control Problems (MOMA HOCP). First, the basic functionality of this technique is validated on the single-vehicle problem of reproducing the Cassini interplanetary cruise.

Interplanetary↗

A Novel Multi-Spacecraft Interplanetary Global Trajectory Optimization Transcription

As the frontier of space exploration continues to advance, so does the design complexity of future interplanetary missions. One avenue of this increasing complexity includes a class of designs known as ``Distributed Spacecraft Missions"; missions where multiple spacecraft coordinate to perform shared objectives. Current approaches for the global trajectory optimization of these Multi-Vehicle Missions (MVMs) are prone to shortcomings including laborious iterative design, considerable human-in-the-loop effort, treatment of the multi-vehicle problem as multiple separate trajectory optimization subproblems (resulting in suboptimal solutions where the whole is less than the sum of its parts), and poor handling of coordination objectives and constraints. There are only a handful of software platforms in existence capable of fully-automated, rapid, interplanetary mission and systems global optimization including the Parallel Global Multiobjective Optimizer (PaGMO), the Gravity Assisted Low-thrust Local Optimization Program (GALLOP), and the Evolutionary Mission Trajectory Generator (EMTG). However, none of these tools is capable of performing such tasks for MVM designs. The work outlined in this paper lays the groundwork for a technique to begin addressing these shortcomings. We present a fully-automated technique which frames interplanetary MVMs as Multi-Objective, Multi-Agent Hybrid Optimal Control Problems (MOMA HOCP). First, the basic functionality of this technique is validated on the single-vehicle problem of reproducing the Cassini interplanetary cruise.

Napier, Sean W.↗

Adaptive Interplanetary Navigation Using Genetic Algorithms

The problem of tuning trajectory determination models for interplanetary navigation is a complex task requiring an intensive search of multiple dynamical and nondynamical models that yield trajectory solutions with minimal errors.

adaptive↗

End-to-End Performance Optimization of a Crewed Lunar Landing Mission Staged from a Near Rectilinear Halo Orbit

A crewed lunar landing mission staged from a Near Rectilinear Halo Orbit (NRHO) presents a unique set of challenges not considered before for the Apollo-type direct missions. The NRHO does not have a fixed orbital plane and its period also changes with time, due to which the mission performance is very sensitive to the timing and location of the NRHO departure and arrival burns for a near-polar surface landing site. For such type of mission staged from the NRHO, an end-to-end optimization approach for the entire mission, from the NRHO departure to surface landing and from surface lift-off to the NRHO insertion, is well-suited for extracting the maximum performance out of this design. In this paper, an integrated approach for performing end-to-end optimization of a NRHO-staged crewed lunar landing mission is proposed that optimizes the in-space and powered flight segments simultaneously using Copernicus trajectory optimization tool. Copernicus is augmented with analytical approximations for the in-space, powered descent, and powered ascent segments in this work to improve the convergence of its multiple-shooting based direct optimization method. New analytical solutions are derived for the different phases of the powered flight segments by solving associated guidance problems. These analytical solutions act as initial guesses for starting the numerical optimization of the integrated mission in Copernicus. An example lunar landing mission is simulated to validate the proposed end-to-end optimization approach and the results are compared with the conventional patched trajectory approach.

Bharat Mahajan↗

GC/MS Method Development for Separating Lunar Volatile Ice Simulant Headspace Gases

Various investigators propose the lunar surface contains widely distributed volatiles, especially water- like species, i.e. OH and H2O. Surface volatiles are theorized to exist as a hydrated regolith layer, concentrated in extremely cold polar permanently shadowed regions (PSR), and/or solar wind implantation reservoirs in lunar glasses. The proposed sources of lunar surface volatiles range from cometary impacts, solar wind, or a supply present during moon formation. Future Artemis missions aim to collect and return the samples containing volatiles collected near lunar polar craters or PSRs. We, as advanced curation scientists, are responsible for developing techniques and methodologies for preserving returned sample integrity as much as possible. Pristine volatile-bearing samples are invaluable to the scientific community seeking to unravel the history of the solar system. Realistically, a sample will experience alteration during collection, transportation back to earth, and storage. The Planetary Exploration and Astromaterials Research Lab (PEARL) seeks to understand temperature and pressure effects on high-fidelity volatile-containing regolith simulants, the foundation for the future of cold curation. This abstract outlines the separation, identification, and quantification of headspace gases over volatile ice feed stock material using gas chromatography/mass spectrometry (GC/MS). Preliminary objectives concentrated on sample handling, reproducibility, and understanding the elution characteristics for each analyte. Initial GC/MS method development experiments utilized diluted static headspace sample preparation. Diluted samples were used because sampling headspace gases directly from a vial containing liquid analyte resulted in overloading of the column and detector. Overloading is evident based on chromatogram peak shapes and instrument contamination, or carry over, between experiments. A mixture of three alcohols were used for a majority of the sample handling and reproducibility studies. Reproducibility was tested via multiple users, calibration curves, and check standards. Stock solutions of condensed lunar volatile analytes included methanol, ammonia in methanol, hydrogen sulfide in water, and an equal volume mixture of methanol, ethanol, and isopropanol. Current samples use room air as the headspace sample matrix, however future experiments will incorporate an inert purge gas, such as argon or nitrogen. Three mL of each analyte solution were capped in separate 20 mL crimp top GC vials. Dilutions were carried out by removing an aliquot of headspace gases with a calibrated 1 mL gastight syringe and immediately transferring to a 20 mL capped crimp top vial. The GC/MS is a Thermo Fisher Trace 1310/ISQ 7000 with a TriPlus RSH autosampler and split/splitless injector module. The experiments outlined in this abstract use the following hardware: a 2.5 mL gastight headspace syringe tool, 1 mm ID x 78.5 mm length ultra-inert straight injection liner, and a TG-BondQ 30 m × 0.32 mm × 10 μm column. Various parameters, such as hardware selection and the temperature, pressure, and split ratio set points, continue to evolve as the overall experiment is refined. Diluted headspace chromatograms were collected for the individual stock solutions. Retention times, peak shapes, and mass spectra were evaluated and added to the data processing method for each molecule of interest. Figure 1 shows the total ion chromatograms for the three major lunar volatile simulant stock solutions: methanol, 7 N ammonia in methanol, and 0.4% hydrogen sulfide in water. Tailing peak shapes for ammonia (2.98 min rt) and water (4.06 min rt) indicate the molecules are not properly eluting from the selected column with the current separation method. Additionally, hydrogen sulfide and ammonia have overlapping peak windows, which could impact quantification. Ongoing experiments aim to address the peak shape and overlapping via the separation method and hardware selection. Sample preparation reproducibility experiments used stock solution containing equal volumes of a non- interactive mixture of methanol, ethanol, and isopropanol. Mass spectrum ion traces were used to identify and quantify all three alcohols. Peaks were automatically detected, identified, and integrated through the mass spectra detection and processing parameters. Calibration response curves and check standards were used to evaluate the validity of the sample preparation procedure. Figure 2 shows the methanol chromatogram peak area versus total headspace dilution volume transferred from the alcohol mixture vial. The calibration response curves and check standards validate sample preparation procedure. Continuing data analysis efforts are working towards correlating the peak area and instrument response factor to the headspace analyte concentration and condensed phase composition. Static headspace gas chromatography theory relies on Dalton’s law, Raoult’s law, Henry’s Law, and the Kolb and Ettre equation to associate peak area to the analyte composition in a non-ideal solution. Equation 1 is a simplified expression derived from the aforementioned theories. Future experiments involve liquid injections of the individual stock solutions, liquid and headspace analysis of various stock solution combinations, and the addition of regolith simulants to the mixtures. Temperature is another variable expected to affect reaction rates and will be explored.

Cecilia L. Amick↗

Multiple Scattering Effects of Trees on L-band Microwave Using a Hybrid Method

The multiple scattering of electromagnetic waves in forests is studied using a two-step hybrid method. First, the T-matrix of a single tree is calculated based on its far-field computed with the full-wave simulations of the FEKO software. The T-matrix captures the multiple scattering caused by the tree structure. Second, the the interactions among different trees are considered using the T-matrices of the individual trees and the Foldy-Lax equations. The result of the two-step hybrid method is validated with FEKO by solving scattering from two trees directly. The multiple scattering effects are illustrated by the field solutions.

Yueh, Simon↗

LTE Electrolyzer Data Collection

The goal for NREL is to collect, develop and publish performance metrics relative to low temperature electrolyzer installations. This will be done through the development of: Secure storage solution to house the collection of data from multiple projects Standardization of data to be collected and analyzed. This will be done using data templates developed with the help of partners involved with electrolyzer installations. Analysis that produces metrics of interest for all stakeholders Aggregation of results from multiple projects to view industry progress as a whole Publication of aggregated results in the form of composite data products (CDPs) Collaboration with Idaho National Lab and their work with high temperature electrolyzer installations will enable efficient use of storage and analysis tools.

data↗

Practical challenges of model predictive control (MPC) for grid interactive small and medium commercial buildings

To the urgent call for mitigating climate change, substantial initiatives have been undertaken to deploy grid-interactive heating, ventilation, and air-conditioning (HVAC) controls, such as model predictive control (MPC) for buildings. These efforts typically aim to curtail peak energy demand, shift load and enhance overall energy efficiency. With the recent development of low-cost MPC technologies that don’t require extensive instrumentation or manual modeling, small and medium commercial buildings (SMCBs), which rarely utilize advanced HVAC control systems, have become candidates for grid-interactive efficient buildings (GEBs). However, despite the potential benefits and maturity of the technology itself, several practical challenges remain in real-world implementation. In this paper, we share the practical challenges that we have encountered in implementing and testing three types of MPC solutions (ON/OFF unit, dualfuel, and VRF systems) on multiple SMCB sites. We describe the MPC deployment process and discuss the lessons learned. The site selection, eligibility, and retrofit availability (e.g., utility price structure, thermostat communications, etc.) are the main discussion points at the beginning of the project. Also, the modeling automation and the best practices for interacting with endusers and handling erroneous situations are presented for successful operations.

woo Ham, Sang↗

Articulated figure positioning by multiple constraints

A problem that arises in positioning an articulated figure is the solution of three-dimensional joint positions (kinematics) when goal positions, rather than joint angles, are given. If more than one such goal is to be achieved, the problem is often solved interactively by positioning or solving one component of the linkage, then adjusting another, then redoing the first, and so on. This iterative process is slow and tedious. The paper presents a method that automatically solves multiple simultaneous joint position goals. The user interface offers a six-degree-of-freedom input device to specify joint angles and goal positions interactively. Examples are used to demonstrate the power and efficiency of this method for key-position animation.

Badler, Norman I.↗

A regression technique for evaluation and quantification for water quality parameters from remote sensing data

The paper attempts to define optical physics and/or environmental conditions under which the linear multiple-regression should be applicable. It is reported that investigation of the signal response shows that the exact solution for a number of optical physics conditions is of the same form as a linearized multiple-regression equation, even if nonlinear contributions from surface reflections, atmospheric constituents, or other water pollutants are included. Limitations on achieving this type of solution are defined. Laboratory data are used to demonstrate that the technique is applicable to water mixtures which contain constituents with both linear and nonlinear radiance gradients. Finally, it is concluded that instrument noise, ground-truth placement, and time lapse between remote sensor overpass and water sample operations are serious barriers to successful use of the technique.

Whitlock, C. H.↗

Integrated Steady-State System Package for Nuclear Thermal Propulsion Analysis Using Multi-Dimensional Thermal Hydraulics and Dimensionless Turbopump Treatment

Nuclear thermal propulsion is an evolving technology that can be utilized for long-distance space travel. This technology yields the advantage of a high thrust and specific impulse, but requires an examination of the potential design adjustments necessary to enhance its feasibility. The development of nuclear thermal propulsion requires a comprehensive understanding of the system-level behavior during transient and steady-state operation. This paper extends our previous research by including the proper handling of turbomachinery with multi-channel thermal hydraulic simulations only for steady-state solutions. The system-level approach presented here enables the treatment of the turbopump components through non-dimensional analysis that eliminates the assumption of constant efficiencies. All the other components within the system (e.g., reflector and core) can be discretized to multiple channels and layers, in which the full thermal hydraulic solution is established. The approach chosen here enables the realistic modeling of the propellant flow within the expander cycle by capturing the pressure losses, mass flow rate splits, and enthalpy gain for various operational conditions. The verification of the package is completed through point comparisons of previous investigations into similar system designs. Furthermore, sensitivity studies are used to benchmark the capabilities of the package and investigate solution variations due to the perturbation of operational conditions and regimes. The sensitivity studies performed here are important to capture variation in flow characteristics (e.g., temperature, pressure, mass flow rates) for different design objectives such as the thrust and specific impulse. This work demonstrates that system-level simulations lacking multi-channel capability and proper turbomachinery treatment may yield higher uncertainties in understanding the engine’s response and characteristics to changing various requirements. This is extremely important when screening the design space of such propulsion systems and when transient simulations are required.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Gross-fragmentation of meteoroids and bulk density of Geminids from photographic fireball records

The explicit solution of the drag and ablation equations of a single nonfragmenting meteoroid moving in any actual atmosphere was published several years ago. The solution yields the theoretical relation of l, the distance flown by the meteoroid in its trajectory, as a function of time, t, assuming that the height, h, is a known function of l. The photographic records of meteors and fireballs are coded by time marks, using a rotating shutter or a similar device to break the moving image. Time is, thus, the independent variable and for each time mark on a meteoroid trajector, the observed distance along the trajectory, l sub obs, as well as the double- or multiple- station photographs of the same meteoroid. Applying this solution to all available Prairie Network (PN) fireball-records, we recognized that the majority of them gave good solutions with standard deviations somewhat bigger than the intrinsic geometrical precision of the data. We also noticed that, on an average, previous methods of evaluation of the meteoroid velocities (interpolation polynomials, numerical differenciation of the observed l sub obs) used up to only several tens of percent of the intrinsic precision of the PN observational data. When residuals of these solutions, i.e. l sub obs - l sub com, were represented as a function of time for about 75 percent of solutions. The remaining 25 percent of residuals showed systematic changes with time exceeding one standard deviation. We tried to explain these systematic time course of residuals by using different meteoroids first computed theoretically and then analyzed by the same model as the natural PN fireballs were. The conclusion of these model computations is that systematic time changes of residuals in the nonfragmenting model exceeding one standard of deviation are caused by sudden gross fragmentation at one or more trajectory points. Thus, we generalized the explicit solution of the drag and ablation equations of a single nonfragmenting meteoroid by allowing for one or more points, where sudden gross fragmentation can occur. Using this generalized solution, the distances along the meteoroid trajectory can be computed for any choice of input parameters and compared with the observed distances flown by the meteoroid. For the most precise and long fireball trajectories, the least-squares solution can, thus, yield the initial velocities, the ablation coefficients, the dynamical masses, the positions of gross-fragmentation points, and the terminal mass. At a gross-fragmentation point, the ratio of the main mass to all the remaining fragments can be compared with the dynamic mass determined from our gross-fragmentation model and, thus, the meteoroid bulk density can be evaluated. We applied the gross-fragmentation model to sever PN fireballs showing time changes of residuals, and we recognized that, in all these cases, the new computed bulk densities of meteoroids resulted higher in comparison with the meteoroid densities determined with the non-gross-fragmentation model. Other aspects of the study are discussed.

Ceplecha, Zdenek↗

Scattering and Diffraction of Electromagnetic Radiation: An Effective Probe to Material Structure

Scattered electromagnetic waves from material bodies of different forms contain, in an intricate way, precise information on the intrinsic, geometrical and physical properties of the objects. Scattering theories, ever deepening, aim to provide dependable interpretation and prediction to the complicated interaction of electromagnetic radiation with matter. There are well-established multiple-scattering formulations based on classical electromagnetic theories. An example is the Generalized Multi-particle Mie-solution (GMM), which has recently been extended to a special version ̶ the GMM-PA approach, applicable to finite periodic arrays consisting of a huge number (e.g., >>106) of identical scattering centers [1]. The framework of the GMM-PA is nearly complete. When the size of the constituent unit scatterers becomes considerably small in comparison with incident wavelength, an appropriate array of such small element volumes may well be a satisfactory representation of a material entity having an arbitrary structure. X-ray diffraction is a powerful characterization tool used in a variety of scientific and technical fields, including material science. A diffraction pattern is nothing more than the spatial distribution of scattered intensity, determined by the distribution of scattering matter by way of its Fourier transform [1]. Since all linear dimensions entered into Maxwell's equations are normalized by wavelength, an analogy exists between optical and X-ray diffraction patterns. A large set of optical diffraction patterns experimentally obtained can be found in the literature [e.g., 2,3]. Theoretical results from the GMM-PA have been scrutinized using a large collection of publically accessible, experimentally obtained Fraunhofer diffraction patterns. As far as characteristic structures of the patterns are concerned, theoretical and experimental results are in uniform agreement; no exception has been found so far. Closely connected with the spatial distribution of scattered intensities are cross sections, such as for extinction, scattering, absorption, and radiation pressure, as a critical type of key quantity addressed in most theoretical and experimental studies of radiative scattering. Cross sections predicted from different scattering theories are supposed to be in general agreement. For objects of irregular shape, the GMM-PA solutions can be compared with the highly flexible Discrete Dipole Approximation (DDA) [4,5] when dividing a target to no more than ~106 unit cells. Also, there are different ways to calculate the cross sections in the GMM-PA, providing an additional means to examine the accuracy of the numerical solutions and to unveil potential issues concerning the theoretical formulations and numerical aspects. To solve multiple scattering by an assembly of material volumes through classical theories such as the GMM-PA, the radiative properties of the component scatterers, the complex refractive index in particular, must be provided as input parameters. When using a PA to characterize a material body, this involves the use of an adequate theoretical tool, an effective medium theory, to connect Maxwell's phenomenogical theory with the atomistic theory of matter. In the atomic theory, one regards matter as composed of interacting particles (atoms and molecules) embedded in the vacuum [6]. However, the radiative properties of atomic-scaled particles are known to be substantially different from bulk materials. Intensive research efforts in the fields of cluster science and nanoscience attempt to bridge the gap between bulk and atom and to understand the transition from classical to quantum physics. The GMM-PA calculations, which place virtually no restriction on the component-particle size, might help to gain certain insight into the transition.

Xu, Yu-Lin↗

UCLA International Conference on Radiation and Remote Probing of the Atmosphere, University of California, Los Angeles, Calif., August 28-30, 1973, Proceedings

Approaches for solving multiple scattering problems in planetary atmospheres are considered along with the solution of the radiative transfer theory problems by the Monte Carlo method, the radiative transfer of visible radiation in turbid atmospheres, aspects of scattering and absorption from poly-dispersed aerosols, and multiple scattering in cloud layers. Other subjects discussed include particulate sizes from polarization measurements, lidar observations of atmospheric particulate content, methods of calculating infrared transfer, molecular absorption parameters in atmospheric modelling, and infrared remote sounding. The influence of the atmosphere on spectral radiance and contrasts of natural formations measured from space is investigated and analytical results for radiative transfer in thick atmospheres are presented. Individual items are announced in this issue.

Kuriyan, J. G.↗

Volume serving and media management in a networked, distributed client/server environment

The E-Systems Modular Automated Storage System (EMASS) is a family of hierarchical mass storage systems providing complete storage/'file space' management. The EMASS volume server provides the flexibility to work with different clients (file servers), different platforms, and different archives with a 'mix and match' capability. The EMASS design considers all file management programs as clients of the volume server system. System storage capacities are tailored to customer needs ranging from small data centers to large central libraries serving multiple users simultaneously. All EMASS hardware is commercial off the shelf (COTS), selected to provide the performance and reliability needed in current and future mass storage solutions. All interfaces use standard commercial protocols and networks suitable to service multiple hosts. EMASS is designed to efficiently store and retrieve in excess of 10,000 terabytes of data. Current clients include CRAY's YMP Model E based Data Migration Facility (DMF), IBM's RS/6000 based Unitree, and CONVEX based EMASS File Server software. The VolSer software provides the capability to accept client or graphical user interface (GUI) commands from the operator's console and translate them to the commands needed to control any configured archive. The VolSer system offers advanced features to enhance media handling and particularly media mounting such as: automated media migration, preferred media placement, drive load leveling, registered MediaClass groupings, and drive pooling.

Herring, Ralph H.↗

Numerical-perturbation technique for stability of flat-plate boundary layers with suction

A numerical-perturbation scheme is proposed for determining the stability of flows over plates with suction through a finite number of porous suction strips. The basic flow is calculated as the sum of the Blasius flow and closed-form linearized triple-deck solutions of the flow due to the strips. A perturbation technique is used to determine the increment a(ij) in the complex wavenumber at a given location x(j) due to the presence of a strip centered at x(i). The end result is a set of influence coefficients that can be used to determine the growth rates and amplification factors for any suction levels without repeating the calculations. The numerical-perturbation results are verified by comparison with interacting boundary layers for the case of six strips and the experimental data of Reynolds and Saric for single- and multiple-strip configurations. The influence coefficient form of the solution suggests a scheme for optimizing the strip configuration. The results show that one should concentrate the suction near branch I of the neutral stability curve, a conclusion verified by the experiments.

Reed, H. L.↗

Fluorescence Studies of Protein Crystal Nucleation

We have postulated that, in the case of tetragonal chicken egg white lysozyme, crystal growth occurs by the addition of pre-critical nuclei sized n-mers that form in the bulk solution, and that the n-mer growth units were multiples of the tetrameric 4(sub 3) helical structure. These have the strongest intermolecular bonds in the crystal and are therefore likely to be the first species formed. High resolution AFM studies provide strong supporting evidence for this model, but the data also suggest that the actual species in solution may not be identical in structure to that found in the crystal. We are using fluorescence resonance energy transfer (FRET) to study the initial solution phase self-assembly process, using covalent fluorescent derivatives which crystallize in the characteristic P4(sub 3)2(sub 1)2(sub 1) space group. FRET studies are being carried out between the cascade blue (CB-lys, donor, Ex(sub max) 366 nm, Em 420 nm) and lucifer yellow (LY-lys, acceptor, Ex(sub max) 430 nm, Em 528 nm) asp101 derivatives. The estimated R(sub 0) for this probe pair, the distance where 50% of the donor energy is transferred to the acceptor, is approx. 1.2 nm, compared to 2.2 nm between the side chain carboxyls of adjacent asp101's in the crystalline 4(sub 3) helix. The short donor lifetime of 2.80 ns (chi(sup 2) = 0.644), coupled with the large average distances between the molecules (greater than or equal to 50 nm) in solution, ensure that any energy transfer observed is not due to random diffusive interactions. Lifetime data show that CB-lys has a single lifetime when it is the only species in solution. Similarly, LY-lys also exhibits a single lifetime of 4.63 ns (chi(sup 2) = 0.42) when alone in solution. Addition of LY-lys to CB-lys results in the appearance of a third lifetime component of 0.348ns for the CB-lys. The fractional intensities of the different species present can be used to estimate the distribution of monomer and n-mers in solution. The self-association process is a function of the protein concentration relative to the saturation concentration, and observing it in dilute solution (conc. less than or equal to 10(exp -5)M) requires that the experiments be performed under low solubility conditions, i.e., low temperatures and high salt concentrations. Data from preliminary steady state FRET studies with N-terminal bound pyrene acetic acid (PAA-lys, donor, Ex 340 nm, Em 376 nm) and asp101 LY-lys as an acceptor showed a consistent trend of decreasing donor fluorescence intensity with increasing total protein concentration. The FRET data have been obtained at pH 4.6, 0.1M NaAc buffer, at 5 and 7% NaCl, 4 C. The corresponding C(sub sat) values are 0.471 and 0.362 mg/ml (approx. 3.3 and approx. 2.5 x 10(exp -5)M respectively). The donor fluorescence decrease is more pronounced at7% NaCl, consistent with the expected increased intermolecular interactions at higher salt concentrations as reflected in the lower solubility. Results from these and other ongoing studies will be discussed in conjunction with an emerging model for how tetragonal lysozyme crystals nucleate and the relevance of that model to other proteins.

Pusey, Marc↗