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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 91 records · Page 5
All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Structural snapshots of base excision by the cancer-associated variant MutY N146S reveal a retaining mechanism

Abstract DNA glycosylase MutY plays a critical role in suppression of mutations resulted from oxidative damage, as highlighted by cancer-association of the human enzyme. MutY requires a highly conserved catalytic Asp residue for excision of adenines misinserted opposite 8-oxo-7,8-dihydroguanine (OG). A nearby Asn residue hydrogen bonds to the catalytic Asp in structures of MutY and its mutation to Ser is an inherited variant in human MUTYH associated with colorectal cancer. We captured structural snapshots of N146S Geobacillus stearothermophilus MutY bound to DNA containing a substrate, a transition state analog and enzyme-catalyzed abasic site products to provide insight into the base excision mechanism of MutY and the role of Asn. Surprisingly, despite the ability of N146S to excise adenine and purine (P) in vitro, albeit at slow rates, N146S-OG:P complex showed a calcium coordinated to the purine base altering its conformation to inhibit hydrolysis. We obtained crystal structures of N146S Gs MutY bound to its abasic site product by removing the calcium from crystals of N146S-OG:P complex to initiate catalysis in crystallo or by crystallization in the absence of calcium. The product structures of N146S feature enzyme-generated β-anomer abasic sites that support a retaining mechanism for MutY-catalyzed base excision.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Identification and Biochemical Characterization of a Novel Hormone-Sensitive Lipase Family Esterase Est19 from the Antarctic Bacterium Pseudomonas sp. E2-15

Esterases represent an important class of enzymes with a wide variety of industrial applications. A novel hormone-sensitive lipase (HSL) family esterase, Est19, from the Antarctic bacterium Pseudomonas sp. E2-15 is identified, cloned, and expressed. The enzyme possesses a GESAG motif containing an active serine (S) located within a highly conserved catalytic triad of Ser 155 , Asp 253 , and His 282 residues. The catalytic efficiency (k cat /K m ) of Est19 for the pNPC6 substrate is 148.68 s –1 mM –1 at 40 °C. Replacing Glu 154 juxtaposed to the critical catalytic serine with Asp (E154→D substitution) reduced the activity and catalytic efficiency of the enzyme two-fold, with little change in the substrate affinity. The wild-type enzyme retained near complete activity over a temperature range of 10–60 °C, while ~50% of its activity was retained at 0 °C. A phylogenetic analysis suggested that Est19 and its homologs may represent a new subfamily of HSL. The thermal stability and stereo-specificity suggest that the Est19 esterase may be useful for cold and chiral catalyses.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2023

Piperidine CD4-Mimetic Compounds Expose Vulnerable Env Epitopes Sensitizing HIV-1-Infected Cells to ADCC

The ability of the HIV-1 accessory proteins Nef and Vpu to decrease CD4 levels contributes to the protection of infected cells from antibody-dependent cellular cytotoxicity (ADCC) by preventing the exposure of Env vulnerable epitopes. Small-molecule CD4 mimetics (CD4mc) based on the indane and piperidine scaffolds such as (+)-BNM-III-170 and (S)-MCG-IV-210 sensitize HIV-1-infected cells to ADCC by exposing CD4-induced (CD4i) epitopes recognized by non-neutralizing antibodies that are abundantly present in plasma from people living with HIV. Here, we characterize a new family of CD4mc, (S)-MCG-IV-210 derivatives, based on the piperidine scaffold which engages the gp120 within the Phe43 cavity by targeting the highly conserved Asp 368 Env residue. We utilized structure-based approaches and developed a series of piperidine analogs with improved activity to inhibit the infection of difficult-to-neutralize tier-2 viruses and sensitize infected cells to ADCC mediated by HIV+ plasma. Moreover, the new analogs formed an H-bond with the α-carboxylic acid group of Asp 368 , opening a new avenue to enlarge the breadth of this family of anti-Env small molecules. Overall, the new structural and biological attributes of these molecules make them good candidates for strategies aimed at the elimination of HIV-1-infected cells.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1972

The study and development of an active scattering particle size spectrometer for space environments

The work is reported on laser operation in a vacuum, and the theoretical response of active scattering particle spectrometers (ASPS). The theoretical considerations connected with the thermal, interferometric and electrical problems that influence laser operation under vacuum conditions are discussed along with the scattering signal pulses with high frequency ripple content which were observed while operating the ASPS.

Knollenberg, R. G.↗
NASA NTRS1977

Digital controller design: Analysis of the annular suspension pointing system

The annular suspension and pointing system (ASPS) a payload auxiliary pointing device of the space shuttle is briefly described along with the function of the digital controller. The equations of motion of a simplified plan planar model of the ASPS are derived. Results of computer simulations are discussed.

Source record↗
NASA NTRS1978

The 1.1 micrometer and visible emission semiconductor diode lasers

In (AlGa)As, the first of three alloy systems studied, Continuous Wave (CW) operation was obtained at room temperature at a wavelength as low as 7260 A. Reliability in this system was studied in the incoherent mode. Zinc doped devices had significant degradation, whereas Ge or Ge plus Zi doped devices had none. The Al2O3 facet coatings were shown to significantly reduce facet deterioration in all types of lasers, longer wavelength units of that type having accumulated (at the time of writing) 22,000 hours with little if any degradation. A CL study of thin (AlGa)As layers revealed micro fluctuation in composition. A macro-scale fluctuation was observed by electroreflectance. An experimental and theoretical study of the effect of stripe width on the threshold current was carried out. Emission below 7000 A was obtained in VPE grown Ga(AsP) (In,Ga)P with CW operation at 10 C. Lasers and LED's were made by LPE in (InGa) (AsP). Laser thresholds of 5 kA/cm2 were obtained, while LED efficiences were on the order of 2%. Incoherent life test over 6000 hours showed no degradation.

Ladany, I.↗
NASA NTRS1978

The Annular Suspension and Pointing System /ASPS/

The Annular Suspension and Pointing System (ASPS) may be attached to a carrier vehicle for orientation, mechanical isolation, and fine pointing purposes applicable to space experiments. It has subassemblies for both coarse and vernier pointing. A fourteen-degree-of-freedom simulation of the ASPS mounted on a Space Shuttle has yielded initial performance data. The simulation describes: the magnetic actuators, payload sensors, coarse gimbal assemblies, control algorithms, rigid body dynamic models of the payload and Shuttle, and a control system firing model.

Anderson, W. W.↗
NASA NTRS1978

The Annular Suspension and Pointing System

The Annular Suspension and Pointing System (ASPS) is a general purpose mount designed to provide orientation, mechanical isolation, and fine pointing for space experiments. The ASPS consists of two assemblies, the first being a set of two gimbals attached to a carrier spacecraft (e.g., Space Shuttle) and providing coarse pointing, and the second a magnetic vernier pointing and isolation assembly attached to the inner gimbal of the first assembly and providing fine pointing. Discussion of the evolution of this concept, required related technology, and data from analyses and simulations predicting pointing accuracies that allowed the specification of hardware design requirements is presented.

Anderson, W. W.↗
NASA NTRS1979

Digital controller design: Analysis of the annular suspension pointing system

The Annular Suspension and Pointing System (ASPS) is a payload auxiliary pointing device of the Space Shuttle. The ASPS is comprised of two major subassemblies, a vernier and a coarse pointing subsystem. The experiment is attached to a mounting plate/rim combination which is suspended on magnetic bearing/actuators (MBA) strategically located about the rim. Fine pointing is achieved by gimballing the plate/rim within the MBA gaps. Control about the experiment line-of-sight is obtained through the use of a non-contacting rim drive and positioning torquer. All sensors used to close the servo loops on the vernier system are noncontacting elements. Therefore, the experiment is a free-flyer constrained only by the magnetic forces generated by the control loops.

Kuo, B. C.↗
NASA NTRS1981

An evaluation of the Interactive Software Invocation System (ISIS) for software development applications

The Interactive Software Invocation System (ISIS), which allows a user to build, modify, control, and process a total flight software system without direct communications with the host computer, is described. This interactive data management system provides the user with a file manager, text editor, a tool invoker, and an Interactive Programming Language (IPL). The basic file design of ISIS is a five level hierarchical structure. The file manager controls this hierarchical file structure and permits the user to create, to save, to access, and to purge pages of information. The text editor is used to manipulate pages of text to be modified and the tool invoker allows the user to communicate with the host computer through a RUN file created by the user. The IPL is based on PASCAL and contains most of the statements found in a high-level programming language. In order to evaluate the effectiveness of the system as applied to a flight project, the collection of software components required to support the Annular Suspension and Pointing System (ASPS) flight project were integrated using ISIS. The ASPS software system and its integration into ISIS is described.

Noland, M. S.↗
NASA NTRS1993

Annular suspension and pointing system with controlled DC electromagnets

The Annular Suspension and Pointing System (ASPS) developed by the Flight System division of Sperry Corporation is a six-degree of freedom payload pointing system designed for use with the space shuttle. This magnetic suspension and pointing system provides precise controlled pointing in six-degrees of freedom, isolation of payload-carrier disturbances, and end mount controlled pointing. Those are great advantages over the traditional mechanical joints for space applications. In this design, we first analyzed the assumed model of the single degree ASPS bearing actuator and obtained the plant dynamics equations. By linearizing the plant dynamics equations, we designed the cascade and feedback compensators such that a stable and satisfied result was obtained. The specified feedback compensator was computer simulated with the nonlinearized plant dynamics equations. The results indicated that an unstable output occurred. In other words, the designed feedback compensator failed. The failure of the design is due to the Taylor's series expansion not converging.

Vu, Josephine Lynn↗
NASA NTRS1996

Design and Implementation of a Digital Controller for a Vibration Isolation and Vernier Pointing System

This paper discusses the recommissioning of the Annular Suspension and Pointing System (ASPS), originally developed in the mid 1970's for pointing and vibration isolation of space experiments. The hardware was developed for NASA Langley Research Center by Sperry Flight Systems (now Honeywell Satellite Systems), was delivered to NASA in 1983. Recently, the hardware was loaned to Old Dominion University (ODU). The ASPS includes coarse gimbal assemblies and a Vernier Pointing Assembly (VPA) that utilize magnetic suspension to provide noncontacting vibration isolation and vernier pointing of the payload. The VPA is the main focus of this research. At ODU, the system has been modified such that it can now be operated in a l-g environment without a gravity offload. Suspension of the annular iron rotor in five degrees-of-freedom has been achieved with the use of modern switching power amplifiers and a digital controller implemented on a 486-class PC.

Neff, Daniel J.↗
NASA NTRS1996

Distribution to the Astronomy Community of the Compressed Digitized Sky Survey

The Space Telescope Science Institute has compressed an all-sky collection of ground-based images and has printed the data on a two volume, 102 CD-ROM disc set. The first part of the survey (containing images of the southern sky) was published in May 1994. The second volume (containing images of the northern sky) was published in January 1995. Software which manages the image retrieval is included with each volume. The Astronomical Society of the Pacific (ASP) is handling the distribution of the lOx compressed data and has sold 310 sets as of October 1996. ASP is also handling the distribution of the recently published 100x version of the northern sky survey which is publicly available at a low cost. The target markets for the 100x compressed data set are the amateur astronomy community, educational institutions, and the general public. During the next year, we plan to publish the first version of a photometric calibration database which will allow users of the compressed sky survey to determine the brightness of stars in the images.

Postman, Marc↗
NASA NTRS1998

Preparation and Characterization of Fluorescent Derivatives of Lysozyme

Fluorescence is one of the most versatile and powerful tools for the study of macromolecules. However, its use in macromolecular crystal growth studies is hampered by the necessity of preparing fluorescent derivatives where the probe does not markedly affect the crystal packing. Alternatively, one can prepare derivatives of limited utility if it is known that they will not affect the specific goals of a given study. We have prepared a number of fluorescent derivatives of chicken egg white lysozyme, covalently attaching fluorescent probes to two different sites on the protein molecule. The first site is the side chain carboxyl group of ASP 101. Amine containing probes such as lucifer yellow, cascade blue, and 5- (2-aminoethyl) aminonapthalene-l-sulfonic acid (EDANS) have been attached using a carbodiimide coupling procedure. ASP 101 lies within the active site cleft, and it is believed that the probes are "buried" within that cleft. This is supported by the fact that all such derivatives have been found to crystallize, with the crystals being fluorescent. Tetragonal crystals of the lucifer yellow derivative have been found to diffract to at least 1.9 A resolution. X-ray diffraction data has been acquired and we are now working on the structure of this derivative. The second group of derivatives is to the N-terminal amine group. The derivatization reaction is performed by using a succinimidyl ester of the probe to be attached. Fluorescent probes such as pyrene acetic acid, 5-carboxyfluorescein, and Oregon green have been attached to this site. We have had little success in crystallizing these derivatives, probably because this site is part of the contact region between the 43 helix chains. However, these sites do not interfere with formation of the 43 helices and the derivatives are suitable for study of their formation in solution. The derivatives are being characterized by steady state and lifetime fluorescence methods, and the presentation will discuss these results.

Smith, Lori↗
NASA NTRS2004

Structural Basis for the Catalytic Activity of Human SER/THR Protein Phosphatase-5

Serinekhreonine protein phosphatase-5 (PP5) affects many signaling networks that regulate cell growth. Here we report the 1.6 Angstrom resolution crystal structure of PP5 catalytic domain with metal and phosphate ions in the active site. The structure reveals a mechanism for PPS-mediated catalysis that requires the precise positioning of two metal ions within a conserved Asp(sup 271)-M(sub 1),-M(sub 2)-His(sup 427)-W(sup 2)-His(sup 304)-Asp(sup 274) catalytic motif, and provides a structural basis for the exceptional catalytic proficiency of protein phosphatases placing them among the most powerful catalysts. Resolution of the entire C-terminus revealed a novel subdomain, and the structure of PP5 should aid development of specific inhibitors.

Swingle, M. R.↗
NASA NTRS2004

Structural Basis for the Catalytic Activity of Human Serine/Threonine Protein Phosphatase type 5 (PP5)

Serine/threonine protein phosphatase-5 (PP5) is a member of the PPP-gene family of protein phosphatases that is widely expressed in mammalian tissues and is highly conserved among eukaryotes. PP5 associates with several proteins that affect signal transduction networks, including the glucocorticoid receptor (GR)-heat shock protein-90 (Hsp90)-heterocomplex, the CDC16 and CDC27 subunits of the anaphase-promoting complex, elF2alpha kinase, the A subunit of PP2A, the G12-alpha / G13-alpha subunits of heterotrimeric G proteins and DNA-PK. The catalytic domain of PP5 (PP5c) shares 35-45% sequence identity with the catalytic domains of other PPP-phosphatases, including protein phosphatase-1 (PP1), -2A (PP2A), -2B / calcineurin (PP2B), -4 (PP4), -6 (PP6), and -7 (PP7). Like PP1, PP2A and PP4, PP5 is also sensitive to inhibition by okadaic acid, microcystin, cantharidin, tautomycin, and calyculin A. Here we report the crystal structure of the PP5 catalytic domain (PP5c) at a resolution of 1.6 angstroms. From this structure we propose a mechanism for PP5-mediated hydrolysis of phosphoprotein substrates, which requires the precise positioning of two metal ions within a conserved Asp(sup 271)-M(sub 1):M(sub 2)-W(sup 1)-His(sup 304)-Asp(sup 274) catalytic motif. The structure of PP5c provides a possible structural basis for explaining the exceptional catalytic proficiency of protein phosphatases, which are among the most powerful known catalysts. Resolution of the entire C-terminus revealed a novel subdomain, and the structure of the PP5c should also aid development of type-specific inhibitors.

Swingle, Mark R.↗
NASA NTRS1992

The back photoreaction of the M intermediate in the photocycle of bacteriorhodopsin: mechanism and evidence for two M species

The back photoreaction of the M intermediate in the photocycle of bacteriorhodopsin is investigated both for the native pigment and its D96N mutant. The experimental setup is based on creating the M intermediate by a first pulse, followed by a (blue) laser pulse which drives the back photoreaction of M. Experiments are carried out varying the delay between the two pulses, as well as the temperature over the -25 degrees C-20 degrees C range. It is found that the kinetic patterns of the M back photoreaction change with time after the generation of this intermediate. The data provide independent evidence for the suggestion of a photocycle mechanism based on two distinct M intermediates. They are thus in keeping with the consecutive model of Varo and Lanyi (Biochemistry 30, 5016-5022; 1991), although they cannot exclude other models such as those based on branched or parallel cycles. More generally, we offer a "photochemical" approach to discriminating between intermediate stages in the photocycle which does not depend on spectroscopic and/or kinetic data. While markedly affecting the rate of the M --> N transition in the photocycle, the rate of the thermal step in back photoreaction of M, at both room and low temperatures, is not significantly affected by the D96N mutation. It is proposed that while Asp 96 is the Schiff-base protonating moiety in the M --> N transition, another residue (most probably Asp 85) reprotonates the Schiff base following light absorption by M.

NASA Discipline Exobiology↗
NASA NTRS1993

Pathways of proton transfer in the light-driven pump bacteriorhodopsin

The mechanism of proton transport in the light-driven pump bacteriorhodopsin is beginning to be understood. Light causes the all-trans to 13-cis isomerization of the retinal chromophore. This sets off a sequential and directed series of transient decreases in the pKa's of a) the retinal Schiff base, b) an extracellular proton release complex which includes asp-85, and c) a cytoplasmic proton uptake complex which includes asp-96. The timing of these pKa changes during the photoreaction cycle causes sequential proton transfers which result in the net movement of a proton across the protein, from the cytoplasmic to the extracellular surface.

NASA Discipline Number 52-30↗