scholarly journals Cloning and high-level expression of monomeric human superoxide dismutase 1 (SOD1) and its interaction with pyrimidine analogs

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0247684
Author(s):  
Marcia LeVatte ◽  
Matthias Lipfert ◽  
Dipankar Roy ◽  
Andriy Kovalenko ◽  
David Scott Wishart
Keyword(s):  
Superoxide Dismutase ◽  
Nmr Spectroscopy ◽  
Nmr Spectra ◽  
Expression System ◽  
Leader Peptide ◽  
Superoxide Dismutase 1 ◽  
H Nmr ◽  
Molecule Interactions ◽  
High Level ◽  
Pyrimidine Analogs

Superoxide dismutase 1 (SOD1) is known to be involved in the pathogenesis of Amyotrophic Lateral Sclerosis (ALS) and is therefore considered to be an important ALS drug target. Identifying potential drug leads that bind to SOD1 and characterizing their interactions by nuclear magnetic resonance (NMR) spectroscopy is complicated by the fact that SOD1 is a homodimer. Creating a monomeric version of SOD1 could alleviate these issues. A specially designed monomeric form of human superoxide dismutase (T2M4SOD1) was cloned into E. coli and its expression significantly enhanced using a number of novel DNA sequence, leader peptide and growth condition optimizations. Uniformly 15N-labeled T2M4SOD1 was prepared from minimal media using 15NH4Cl as the 15N source. The T2M4SOD1 monomer (both 15N labeled and unlabeled) was correctly folded as confirmed by 1H-NMR spectroscopy and active as confirmed by an in-gel enzymatic assay. To demonstrate the utility of this new SOD1 expression system for NMR-based drug screening, eight pyrimidine compounds were tested for binding to T2M4SOD1 by monitoring changes in their 1H NMR and/or 19F-NMR spectra. Weak binding to 5-fluorouridine (FUrd) was observed via line broadening, but very minimal spectral changes were seen with uridine, 5-bromouridine or trifluridine. On the other hand, 1H-NMR spectra of T2M4SOD1 with uracil or three halogenated derivatives of uracil changed dramatically suggesting that the pyrimidine moiety is the crucial binding component of FUrd. Interestingly, no change in tryptophan 32 (Trp32), the putative receptor for FUrd, was detected in the 15N-NMR spectra of 15N-T2M4SOD1 when mixed with these uracil analogs. Molecular docking and molecular dynamic (MD) studies indicate that interaction with Trp32 of SOD1 is predicted to be weak and that there was hydrogen bonding with the nearby aspartate (Asp96), potentiating the Trp32-uracil interaction. These studies demonstrate that monomeric T2M4SOD1 can be readily used to explore small molecule interactions via NMR.

Combining in-cell NMR and X-ray fluorescence microscopy to reveal the intracellular maturation states of human superoxide dismutase 1

2015 ◽  
Vol 51 (3) ◽  
pp. 584-587 ◽  
Author(s):  
E. Luchinat ◽  
A. Gianoncelli ◽  
T. Mello ◽  
A. Galli ◽  
L. Banci
Keyword(s):  
Superoxide Dismutase ◽  
Nmr Spectroscopy ◽  
Fluorescence Microscopy ◽  
Superoxide Dismutase 1 ◽  
X Ray ◽  
Optical Fluorescence ◽  
Human Sod1

Combined in-cell NMR spectroscopy, X-ray fluorescence and optical fluorescence microscopies allow describing the intracellular maturation states of human SOD1.

Magnetic resonance imaging (1H) and spectroscopy (1H, 31P) of growing chicken embryos

1993 ◽  
Vol 73 (4) ◽  
pp. 953-965 ◽  
Author(s):  
A. Lirette ◽  
Z. Liu ◽  
D. C. Crober ◽  
R. A. Towner ◽  
U. M. Oehler ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Magnetic Resonance ◽  
Nmr Spectra ◽  
Surface Coil ◽  
Nmr Imaging ◽  
Chicken Embryos ◽  
H Nmr ◽  
Imaging And Spectroscopy ◽  
Two Peaks

Nuclear magnetic resonance (NMR) imaging and spectroscopy techniques were used to observe in vivo anatomical and metabolite changes, respectively, in developing chicken embryos. Proton (1H) NMR images of the eggs revealed major changes in yolk shape from day 2 to day 6. Embryos were visible from day 6 to hatching, and good embryonic anatomical images were obtained. Two peaks were observed from 1H-NMR spectroscopy of fertilized eggs: one for lipid methylene protons, and one for water protons. Water peak to lipid peak ratios did not vary significantly (P > 0.05) from day 2 to day 21 of incubation. Localized 31P-NMR spectra of developing embryos were obtained with either a 31P surface coil or a double-tuned 31P/1H volume coil. The surface-coil method gave a greater signal to noise ratio by a factor of four. The 31P-NMR spectra indicated two peaks at day 2; these were attributed to phosphomonoesters and phosphodiesters. The three peaks characteristic of ATP appeared on day 11 and increased in size until hatching. From day 19, phosphocreatine was detectable. There appeared to be a good correlation between 31P-metabolite changes detected by in vivo 31P-NMR spectroscopy and literature values for biochemical analyses of developing chicken embryos. The advantage in using NMR imaging and spectroscopy techniques is that anatomical and metabolic changes can be obtained in vivo, non-invasively and repeatedly as an embryo develops. Key words: NMR, MRI, embryo, poultry

scholarly journals Assessment of 1H NMR Spectroscopy for Specific Metabolite Fingerprinting of Angelica sinensis

2008 ◽  
Vol 3 (5) ◽  
pp. 1934578X0800300
Author(s):  
Dongmin Su ◽  
Jinglan Han ◽  
Shishan Yu ◽  
Hailin Qin
Keyword(s):  
Nmr Spectroscopy ◽  
1H Nmr ◽  
1H Nmr Spectroscopy ◽  
Nmr Spectra ◽  
Authentic Sample ◽  
Angelica Sinensis ◽  
Metabolite Fingerprinting ◽  
H Nmr ◽  
Pure Compounds ◽  
Polar Extracts

The 1H NMR fingerprints of fractionated non-polar extracts (CSPD A) from the roots of Angelica sinensis of six different specimens were assigned by comparison with the 1H NMR spectra of the isolated pure compounds. The 1H NMR fingerprints showed exclusively characteristic resonance signals of the major constituents of the plant. The 1H NMR fingerprint established for an authentic sample of A. sinensis can be used for authenticating A. sinensis species.

Morpholines-(IV)

1971 ◽  
Vol 26 (11) ◽  
pp. 1140-1143 ◽  
Author(s):  
Riaz F. Abdulla ◽  
Alok N. Bannerji
Keyword(s):  
Nmr Spectroscopy ◽  
Nmr Spectra ◽  
Beta Lactams ◽  
Aryl Substituent ◽  
Open Chain ◽  
H Nmr

Cyclodehydrohalogenation-beta-Lactams-Morpholin-3-ones-H'-NMR SpectroscopyN-Aryl-N-chloroacetyl-2-chlorophenacylamines (2) give morpholinones 3, or beta-lactams 4, depending upon the N-aryl-substituent. N-Phenyl-N- (2,3-dibromo-3-phenylpropionyl) -4-nitrophenacylamine did not undergo base-catalysed cyclization but gave, instead, the α,β-unsaturated, open-chain amide 5. N-Aryl-N-chloroacetyl-aminomethyl-2-naphthylketones afford only beta-lactams. The first attempt at the synthesis of a 2-chloro-3-oxo-morpholine resulted in the isolation of the 2-hydroxyderivative. The H1-NMR spectra of some more 3-oxo-morpholines have been recorded.

Multiaxial Reorientationsof ND+4 Ions Studied by 2H-NMR Spectroscopy

1991 ◽  
Vol 46 (10) ◽  
pp. 829-840 ◽  
Author(s):  
Z. T. Lalowicz ◽  
S. F. Sagnowski
Keyword(s):  
Nmr Spectroscopy ◽  
Density Matrix ◽  
Nmr Spectra ◽  
Activation Energies ◽  
Matrix Formalism ◽  
2H Nmr ◽  
Jump Frequency ◽  
H Nmr ◽  
2H Nmr Spectroscopy ◽  
Density Matrix Formalism

AbstractThe density matrix formalism is used to simulate motional averaging in the 2H-NMR spectra of reorienting ND+4 ions. The development of the spectra under increasing jump frequency about a single C3 or C2 axis is followed. Next we assume a hierarchy of axes in terms of activation energies sufficient to reach extreme narrowing conditions for some axes before activating jumps about a next one. Primary reorientations about the fastest C3 or C2 symmetry axes define the shape of spectra, the width of which is then stepwise reduced by fast reorientations about the subsequent axes in a postulated sequence of statistically uncorrelated jumps.

Light induced E–Z isomerization in a multi-responsive organogel: elucidation from 1H NMR spectroscopy

2015 ◽  
Vol 51 (53) ◽  
pp. 10680-10683 ◽  
Author(s):  
Sanjoy Mondal ◽  
Priyadarshi Chakraborty ◽  
Partha Bairi ◽  
Dhruba P. Chatterjee ◽  
Arun K. Nandi
Keyword(s):  
Mechanical Properties ◽  
Nmr Spectroscopy ◽  
1H Nmr ◽  
1H Nmr Spectroscopy ◽  
Nmr Spectra ◽  
H Nmr ◽  
Imine Bond

Light induced E–Z isomerization along imine bond in a multiresponsive organogel of anthracene attached 3,4,5-tris(dodecyloxy)benzohydrazide gelator altering morphology, fluorescence and mechanical properties is elucidated from 1H NMR spectra.

Oligophosphan-Liganden, X II [1] Pseudohalogeno- und Halogenohydridoeisen(II)-Komplexe des tripodal-tetratertiären Phosphanliganden P(CH2CH2CH2PMe2)3 / Oligophosphine Ligands, X II [1] Pseudohalo and Halohydro Complexes of Iron (II) Containing the Tripod Tetratertiary Phosphine Ligand P(CH2CH2CH2PMe2)3

1985 ◽  
Vol 40 (11) ◽  
pp. 1485-1489 ◽  
Author(s):  
Martin Antberg ◽  
Lutz Dahlenburg
Keyword(s):  
Nmr Spectroscopy ◽  
Nmr Spectra ◽  
Phosphine Ligand ◽  
Octahedral Coordination ◽  
Coordination Geometry ◽  
H Nmr

The iron(II) complexes FeX2[P(CH2CH2CH2PMe2)3] with X = CN (4), N3 (5). and NCS (6) have been prepared by metathetical reaction of FeCl2[P(CH2CH2CH2PMe2)3] (1) with the corresponding sodium pseudohalide. Treatment of the dihalo compounds FeX2[P(CH2CH2CH2PMe2)3] (X2 = Cl2 (1). Br2 (2), I2 (3)) with LiAlH4 gave the halohydro derivatives FeHX[P(CH2CH2CH2PMe2)3], where HX = HCl (7). HBr (8). and HI (9). Consistent with a cis-octahedral coordination geometry, the 31P NMR spectra of 1−9 exhibit AMX2 type splitting. The halohydro complexes 7−9 have also been studied by IR and 1H NMR spectroscopy. 57Fe Möβbauer data have been collected for compounds 1, 4, and 5.

Some reactions of trimeric silicon phthalocyanine under alkaline conditions

2003 ◽  
Vol 07 (03) ◽  
pp. 176-180 ◽  
Author(s):  
Viktor N. Myakov ◽  
Vyacheslav A. Kuropatov ◽  
Tatyana I. Lopatina ◽  
Valentina N. Sedelnikova
Keyword(s):  
Nmr Spectroscopy ◽  
Spectroscopic Study ◽  
Nmr Spectra ◽  
Ring Current ◽  
Electron System ◽  
Absorption Bands ◽  
H Nmr ◽  
Alkaline Conditions ◽  
Silicon Phthalocyanine

The reaction of bis(trimethylsiloxy)triphthalocyaninatosilicon, Me 3 Si ( OSiPc )3 OSiMe 3 ( Pc 3) with KOH in o-xylene in the presence of 15-crown-5 in vacuum at 200°C has been studied by means of UV-vis, EPR and 1 H NMR spectroscopy. Upon gradual removal of H 2 O , the formation of mono- and dianions of Pc 3 with absorption bands positioned at 575-590 nm was observed. An EPR spectroscopic study revealed that both monoanionic and dianionic of Pc 3 are paramagnetic in solution. 1 H NMR spectra have shown the absence of a ring current in the π-electron system of dianion. The reaction of Pc 3 with KOH or metal Na led to degradation of the Si - O - Si - backbone happened.

Conformational study of blocked pentofuranoses and their 1,5-anhydro derivatives by 1H NMR spectroscopy

1984 ◽  
Vol 49 (1) ◽  
pp. 235-243
Author(s):  
Jaroslav Zajíček ◽  
Miloš Buděšínský ◽  
Jiří Farkaš
Keyword(s):  
Spectral Analysis ◽  
Nmr Spectroscopy ◽  
Quantum Chemical Calculations ◽  
1H Nmr ◽  
Quantum Chemical ◽  
1H Nmr Spectroscopy ◽  
Nmr Spectra ◽  
Conformational Study ◽  
H Nmr

Preferred conformations of the furanose ring and conformer population about the C(5)-C(4) bond in a series of benzoylated pentofuranosyl cyanides have been determined from their 1H NMR spectra. For the 1,5-anhydropentofuranose derivatives the results of the 1H NMR spectral analysis have been compared with the quantum chemical calculations.

Some reactions of dimeric silicon phthalocyanines under alkaline conditions

2002 ◽  
Vol 06 (05) ◽  
pp. 336-339 ◽  
Author(s):  
Viktor N. Myakov ◽  
M. A. Lopatin ◽  
Yu. A. Kurskii
Keyword(s):  
Nmr Spectroscopy ◽  
Crown Ether ◽  
Nmr Spectra ◽  
Ring Current ◽  
Electron System ◽  
H Nmr ◽  
Silicon Phthalocyanines ◽  
Alkaline Conditions

Reactions of bis(hydroxy)- and bis(trimethylsiloxy)diphthalocyaninato silicon with KOH were studied in o-xylene or oligosiloxane in the presence of crown ether 15-cr-5 in vacuo at 190-200 °C by the means of electronic, EPR and 1 H NMR spectroscopy. Under gradual removing of H 2 O , consecutive formation of paramagnetic silicon diphthalocyanine monoanions (λ max 580 nm ), and diamagnetic silicon diphthalocyanine dianions (λ max 550 nm ) have been observed. The dianions react with H 2 O to regenerate silicon diphthalocyanine monoanions and then the starting silicon diphthalocyanine. Reaction of the dianion with O 2 results in a decomposition of silicon diphthalocyanine. 1 H NMR spectra of silicon diphthalocyanine dianion have proven the absence of a ring current in the π-electron system of the dianion.

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