scholarly journals NMR Characterization of Angiogenin Variants and tRNAAla Products Impacting Aberrant Protein Oligomerization

Author(s):  
Andrea Fagagnini ◽  
Sabrina Fasoli ◽  
Miguel Garavís ◽  
Irene Gómez-Pinto ◽  
Giovanni Gotte ◽  
...  

Protein oligomerzation is key to countless physiological processes, but also to abnormal amyloid conformations implicated in over 25 mortal human diseases. Angiogenin (h-ANG), a ribonuclease A family member, produces RNA fragments that regulate ribosome formation, the creation of new blood vessels and stress granule function. Too little h-ANG activity leads abnormal protein oligomerization resulting in Amyotrophic Lateral Sclerosis (ALS) or Parkinson’s disease. While a score of disease linked h-ANG mutants has been studied by X-ray diffraction, some elude crystallization. There is also debate regarding the structure that RNA fragments adopt after cleavage by h-ANG. Here, to better understand the beginning of the process that leads to aberrant protein oligomerization, the solution secondary structure and residue-level dynamics of WT h-ANG and two mutants: H13A and R121C, are characterized by multidimensional heteronuclear NMR spectroscopy under near physiological conditions. All three variants are found to adopt well folded and highly rigid structures in solution, although the elements of secondary structure are somewhat shorter than those observed in cystallography studies. R121C alters the environment of nearby residues only. By constrast, the mutation H13A affects local residues as well as nearby active site residues residues K41 and H119. The conformation characterization by CD and 1D 1H NMR spectroscopies of tRNAAla before and after h-ANG cleavage reveals a retention of most duplex structure and little or no G-quadruplex formation.

2021 ◽  
Vol 22 (3) ◽  
pp. 1439
Author(s):  
Andrea Fagagnini ◽  
Miguel Garavís ◽  
Irene Gómez-Pinto ◽  
Sabrina Fasoli ◽  
Giovanni Gotte ◽  
...  

Protein oligomerization is key to countless physiological processes, but also to abnormal amyloid conformations implicated in over 25 mortal human diseases. Human Angiogenin (h-ANG), a ribonuclease A family member, produces RNA fragments that regulate ribosome formation, the creation of new blood vessels and stress granule function. Too little h-ANG activity leads to abnormal protein oligomerization, resulting in Amyotrophic Lateral Sclerosis (ALS) or Parkinson’s disease. While a score of disease linked h-ANG mutants has been studied by X-ray diffraction, some elude crystallization. There is also a debate regarding the structure that RNA fragments adopt after cleavage by h-ANG. Here, to better understand the beginning of the process that leads to aberrant protein oligomerization, the solution secondary structure and residue-level dynamics of WT h-ANG and two mutants i.e., H13A and R121C, are characterized by multidimensional heteronuclear NMR spectroscopy under near-physiological conditions. All three variants are found to adopt well folded and highly rigid structures in the solution, although the elements of secondary structure are somewhat shorter than those observed in crystallography studies. R121C alters the environment of nearby residues only. By contrast, the mutation H13A affects local residues as well as nearby active site residues K40 and H114. The conformation characterization by CD and 1D 1H NMR spectroscopies of tRNAAla before and after h-ANG cleavage reveals a retention of the duplex structure and little or no G-quadruplex formation.


2021 ◽  
Vol 22 (24) ◽  
pp. 13403
Author(s):  
Sara Saber Younes Mohamed ◽  
Sonia Martinez ◽  
Mauro Banchero ◽  
Luigi Manna ◽  
Silvia Ronchetti ◽  
...  

In the context of the development of carriers for amino acids delivery, Spherical Mesoporous Silica Particles (SMSP), characterized by particles size ranging from 0.15 µm to 0.80 µm and average pore diameter of 2.4 nm, were synthesised and loaded with L-arginine (ARG), a basic amino acid involved in several physiological processes. The loading was performed using water as a solvent through the wet impregnation method (with a final arginine content of 9.1% w/w). The material was characterized before and after impregnation by means of X-Ray Diffraction (XRD), nitrogen sorption analysis, Field Emission Scanning Electron Microscopy (FESEM) and Fourier Transform Infrared (FT-IR) spectroscopy. SMSP are shown to suffer degradation upon impregnation, which dramatically affects their porosity. To elucidate the role of the pH of the ARG impregnating solution (originally set at pH ≈ 11) on SMSP degradation, the loading was performed under different pH conditions (5 and 9) keeping constant the ARG concentration. The impregnation performed with acidic solution did not modify the carrier. All samples displayed ARG in amorphous form: zwitterionic species were present in SMSP impregnated at basic pH whereas positive protonated species in that impregnated at acidic pH.


Author(s):  
T. Gulik-Krzywicki ◽  
M.J. Costello

Freeze-etching electron microscopy is currently one of the best methods for studying molecular organization of biological materials. Its application, however, is still limited by our imprecise knowledge about the perturbations of the original organization which may occur during quenching and fracturing of the samples and during the replication of fractured surfaces. Although it is well known that the preservation of the molecular organization of biological materials is critically dependent on the rate of freezing of the samples, little information is presently available concerning the nature and the extent of freezing-rate dependent perturbations of the original organizations. In order to obtain this information, we have developed a method based on the comparison of x-ray diffraction patterns of samples before and after freezing, prior to fracturing and replication.Our experimental set-up is shown in Fig. 1. The sample to be quenched is placed on its holder which is then mounted on a small metal holder (O) fixed on a glass capillary (p), whose position is controlled by a micromanipulator.


2017 ◽  
Vol 13 (2) ◽  
pp. 4640-4647
Author(s):  
A. M. Abdelghany ◽  
M.S. Meikhail ◽  
S.I. Badr ◽  
A. S. Momen

Thin film samples of pristine polyvinyl chloride (PVC), poly vinyldine fluoride (PVDF) in combination with their blend in addition to samples containing factorial mass fraction of multi wall carbon nano-tubes (MWCNTs) in the dopant level were prepared via routine casting technique using tetrahydrofurane (THF) as a common solvent. X-ray diffraction and transmission electron microscopy (TEM) depict the nano-scale (15-25 nm) of functionalized MWCNTs with no surface damage results from functionalization process.X-ray diffraction (XRD) shows a semi-crystalline nature of PVDF with evidence for more than one phase namely a and b phases. The fraction of b phase was calculated and correlated to the dopant content. FTIR optical absorption spectra revels a preservation of the main vibrational bands before and after addition of MWCNTs in the doping level with a presence of new small band 1151 cm-1 assigned for the interaction and complexation between constituents.


2011 ◽  
Vol 314-316 ◽  
pp. 273-278
Author(s):  
Yu Hua Dong ◽  
Ke Ren ◽  
Qiong Zhou

Linear low density polyethylene (LLDPE) was chemically modified with grafting maleic anhydride (MAH) monomer on its backbone by melting blending. Nano-particles SiO2 was modified by cationic surfactant hexadecyl trimethyl ammonium bromide (CTAB) and anionic surfactant sulfosalicylic acid (SSA) and added to PE coating respectively. Measurement of membrane potential showed that the coating containing modified SiO2 nano-particles had characteristic of ion selectivity. The properties of the different coatings were investigated according to relative industrial standards. Experimental results indicated that PE coating with ion selectivity had better performances, such as adhesion strength, cathodic disbonding and anti-corrosion, than those of coating without ion selectivity. Crystal structure of the coatings before and after alkali corrosion was characterized by Fourier transform infrared spectra (FTIR) and X-ray diffraction (XRD). Structure of the coating without ion selectivity was damaged by NaOH alkali solution, causing mechanical properties being decreased. And the structure of the ion selective coatings was not affected.


2021 ◽  
Vol 22 (2) ◽  
pp. 749
Author(s):  
Patricia B. Gratal ◽  
Julia G. Quero ◽  
Adrián Pérez-Redondo ◽  
Zoila Gándara ◽  
Lourdes Gude

A novel quadruplex ligand based on 1,10-phenanthroline and incorporating two guanyl hydrazone functionalities, PhenQE8, is reported herein. Synthetic access was gained in a two-step procedure with an overall yield of 61%. X-ray diffraction studies revealed that PhenQE8 can adopt an extended conformation that may be optimal to favor recognition of quadruplex DNA. DNA interactions with polymorphic G-quadruplex telomeric structures were studied by different techniques, such as Fluorescence resonance energy transfer (FRET) DNA melting assays, circular dichroism and equilibrium dialysis. Our results reveal that the novel ligand PhenQE8 can efficiently recognize the hybrid quadruplex structures of the human telomeric DNA, with high binding affinity and quadruplex/duplex selectivity. Moreover, the compound shows significant cytotoxic activity against a selected panel of cultured tumor cells (PC-3, HeLa and MCF-7), whereas its cytotoxicity is considerably lower in healthy human cells (HFF-1 and RPWE-1).


Micromachines ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 643
Author(s):  
Xuhang Zhou ◽  
Qiulin Tan ◽  
Xiaorui Liang ◽  
Baimao Lin ◽  
Tao Guo ◽  
...  

Performing high-temperature measurements on the rotating parts of aero-engine systems requires wireless passive sensors. Surface acoustic wave (SAW) sensors can measure high temperatures wirelessly, making them ideal for extreme situations where wired sensors are not applicable. This study reports a new SAW temperature sensor based on a langasite (LGS) substrate that can perform measurements in environments with temperatures as high as 1300 °C. The Pt electrode and LGS substrate were protected by an AlN passivation layer deposited via a pulsed laser, thereby improving the crystallization quality of the Pt film, with the function and stability of the SAW device guaranteed at 1100 °C. The linear relationship between the resonant frequency and temperature is verified by various high-temperature radio-frequency (RF) tests. Changes in sample microstructure before and after high-temperature exposure are analyzed using scanning electron microscopy (SEM) and X-ray diffraction (XRD). The analysis confirms that the proposed AlN/Pt/Cr thin-film electrode has great application potential in high-temperature SAW sensors.


Author(s):  
F. Mostefa ◽  
Nasr Eddine Bouhamou ◽  
H.A. Mesbah ◽  
Salima Aggoun ◽  
D. Mekhatria

This work aims to study the feasibility of making a geopolymer cement based on dredged sediments, from the Fergoug dam (Algeria) and to evaluate their construction potential particularly interesting in the field of special cementitious materials. These sediments due to their mineralogical composition as aluminosilicates; are materials that can be used after heat treatment. Sedimentary clays were characterized before and after calcination by X-ray diffraction, ATG / ATD, spectroscopy (FTIR) and XRF analysis. The calcination was carried out on the raw material sieved at 80 μm for a temperature of 750 ° C, for 3.4 and 5 hours. The reactivity of the calcined products was measured using isothermal calorimetric analysis (DSC) on pastes prepared by mixing an alkaline solution of sodium hydroxide (NaOH) 8 M in an amount allowing to have a Na / Al ratio close to 1 (1: 1). Also, cubic mortar samples were prepared with a ratio L / S: 0.8, sealed and cured for 24 hours at 60 ° C and then at room temperature until the day they were submited to mechanical testing. to check the extent of geopolymerization. The results obtained allowed to optimize the calcination time of 5 hours for a better reactivity of these sediments, and a concentration of 8M of sodium hydroxide and more suitable to have the best mechanical performances.


2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Kooliyankal Naseema ◽  
Kaniyamkandy Ribin ◽  
Nidiyanga Navya ◽  
Prasoon Prasannan

AbstractNano crystalline zinc sulfide thin films were deposited onto glass substrates by chemical bath deposition method. One of the samples was annealed at 300 °C for 2 h in air using a muffle furnace. The prepared thin films were investigated by X-ray diffraction (XRD), UV–visible spectroscopy (UV–vis), photoluminescence spectroscopy (PL), scanning electron microscopy (SEM) and Raman spectroscopy (FT-R) studies before and after annealing. The analysis confirmed the thermal-induced anion substitution and conversion of ZnS crystal to ZnO wurtzite crystal. XRD pattern showed that these films were phase pure and polycrystalline in nature. Optical band gap was found to be 3.86 eV for ZnS and 3.21 eV for ZnO. The films prepared by this simple, low-cost technique are suitable for photovoltaic and optoelectronic applications.


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