Gene/Protein
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Target Concepts:
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Query: EC:4.1.2.42 (
DTA
)
1,693
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Diphtheria toxin (DT) has been studied as a model for understanding active-site structure and function in the ADP-ribosyltransferases. Earlier evidence suggested that histidine-21 of DT is important for the ADP-ribosylation of eukaryotic elongation factor 2 (EF-2). We have generated substitutions of this residue by cassette mutagenesis of a synthetic gene encoding the catalytic A fragment (
DTA
) of DT, and have characterized purified mutant forms of this domain. Changing histidine-21 to alanine, aspartic acid, leucine, glutamine, or
arginine
diminished ADP-ribosylation activity by 70-fold or greater. In contrast, asparagine proved to be a functionally conservative substitution, which reduced ADP-ribosylation activity by < 3-fold. The asparagine mutant was approximately 50-fold-attenuated in NAD glycohydrolase activity, however. Dissociation constants (Kd) for NAD binding, determined by quenching of the intrinsic protein fluorescence, were 15 microM for wild-type
DTA
, 160 microM for the asparagine mutant, and greater than 500 microM NAD for the alanine, leucine, glutamine, and
arginine
mutants. These and previous results support a model of the ADP-ribosylation of EF-2 in which histidine-21 serves primarily a hydrogen-bonding function. We propose that the pi-imidazole nitrogen of His-21 hydrogen-bonds to the nicotinamide carboxamide, orienting the N-glycosidic bond of NAD for attack by the incoming nucleophile in a direct displacement mechanism, and then stabilizing the transition-state intermediate of this reaction.
...
PMID:Active-site mutations of the diphtheria toxin catalytic domain: role of histidine-21 in nicotinamide adenine dinucleotide binding and ADP-ribosylation of elongation factor 2. 817 90
The lethal factor (LF) and edema factor (EF) of anthrax toxin bind by means of their amino-terminal domains to protective antigen (PA) on the surface of toxin-sensitive cells and are translocated to the cytosol, where they act on intracellular targets. Genetically fusing the amino-terminal domain of LF (LFN; residues 1-255) to certain heterologous proteins has been shown to potentiate these proteins for PA-dependent delivery to the cytosol. We report here that short tracts of lysine,
arginine
, or histidine residues can also potentiate a protein for such PA-dependent delivery. Fusion of these polycationic tracts to the amino terminus of the enzymic A chain of diphtheria toxin (
DTA
; residues 1-193) enabled it to be translocated to the cytosol by PA and inhibit protein synthesis. The efficiency of translocation was dependent on tract length: (LFN > Lys8 > Lys6 > Lys3). Lys6 was approximately 100-fold more active than Arg6 or His6, whereas Glu6 and (SerSerGly)2 were inactive. Arg6DTA was partially degraded in cell culture, which may explain its low activity relative to that of Lys6DTA. The polycationic tracts may bind to anionic sites at the cell surface (possibly on PA), allowing the fusion proteins to be coendocytosed with PA and delivered to the endosome, where translocation to the cytosol occurs. Excess free LFN blocked the action of LFNDTA, but not of Lys6DTA. This implies that binding to the LF/EF site is not an obligatory step in translocation and suggests that the polycationic tag binds to a different site. Besides elucidating the process of translocation in anthrax toxin, these findings may aid in developing systems to deliver heterologous proteins and peptides to the cytoplasm of mammalian cells.
...
PMID:Fused polycationic peptide mediates delivery of diphtheria toxin A chain to the cytosol in the presence of anthrax protective antigen. 871 Aug 89
L-Arginine
tetrafluoroborate (C6H15N4(+)O2BF4(-)) is a semiorganic crystalline material exhibiting second harmonic generation (SHG) down to the UV region. The material was synthesized and characterized by elemental analysis, FT-NMR and FT-IR spectra. The lower cutoff wavelength was found to be 270 nm, with a good optical transmittance window covering UV-vis-NIR region. The thermal stability of the compound was determined by TG-
DTA
curves. The powder SHG efficiency was measured using a Q-Switched Nd:YAG laser assembly by a modified Kurtz and Perry powder technique and its phase matching property was established.
...
PMID:Phase matchable semiorganic NLO material for frequency doubling: L-Arginine tetrafluoroborate. 1634 44
Single crystals of L-
arginine
maleate dihydrate (LAMD) were successfully grown from aqueous solution by solvent evaporation technique. As-grown crystals were analyzed by different instrumentation techniques such as X-ray powder diffraction (XRD), Fourier transform infrared (FTIR) spectra and UV-vis near infrared (NIR) transmittance spectra. Thermal behavior has been studied with TGA/
DTA
analyses. The optical second harmonic generation (SHG) conversion efficiency of LAMD was determined using Kurtz powder technique and found to be 1.5 times that of KDP.
...
PMID:Growth and characterization of NLO based L-arginine maleate dihydrate single crystal. 1991 62
Single crystals of l-
arginine
4-nitrophenolate 4-nitrophenol dihydrate (LAPP) have been grown successfully from the solution of l-
arginine
and 4-nitrophenol. Slow evaporation of solvent technique was adopted to grow the bulk single crystals. Single crystal X-ray diffraction analysis confirms the grown crystal has monoclinic crystal system with space group of P21. Powder X-ray diffraction analysis shows the good crystalline nature. The crystalline perfection of the grown single crystals was analyzed by HRXRD by employing a multicrystal X-ray diffractometer. The functional groups were identified from proton NMR spectroscopic analysis. Linear and nonlinear optical properties were determined by UV-Vis spectrophotometer and Kurtz powder technique respectively. It is found that the grown crystal has no absorption in the green wavelength region and the SHG efficiency was found to be 2.66 times that of the standard KDP. The Thermal stability of the crystal was found by obtaining TG/
DTA
curve. The mechanical behavior of the grown crystal has been studied by Vicker's microhardness method.
...
PMID:Crystal growth, structural, thermal and mechanical behavior of l-arginine 4-nitrophenolate 4-nitrophenol dihydrate (LAPP) single crystals. 2496 45