scholarly journals Insights into the molecular determinants of thermal stability in halohydrin dehalogenase HheD2

FEBS Journal ◽  
2021 ◽  
Author(s):  
Julia Wessel ◽  
Giovanna Petrillo ◽  
Miquel Estevez‐Gay ◽  
Sandra Bosch ◽  
Margarita Seeger ◽  
...  

2021 ◽  
Vol 22 (11) ◽  
pp. 5961
Author(s):  
Jelena Vucinic ◽  
Gleb Novikov ◽  
Cédric Y. Montanier ◽  
Claire Dumon ◽  
Thomas Schiex ◽  
...  

With the growing need for renewable sources of energy, the interest for enzymes capable of biomass degradation has been increasing. In this paper, we consider two different xylanases from the GH-11 family: the particularly active GH-11 xylanase from Neocallimastix patriciarum, NpXyn11A, and the hyper-thermostable mutant of the environmentally isolated GH-11 xylanase, EvXyn11TS. Our aim is to identify the molecular determinants underlying the enhanced capacities of these two enzymes to ultimately graft the abilities of one on the other. Molecular dynamics simulations of the respective free-enzymes and enzyme–xylohexaose complexes were carried out at temperatures of 300, 340, and 500 K. An in-depth analysis of these MD simulations showed how differences in dynamics influence the activity and stability of these two enzymes and allowed us to study and understand in greater depth the molecular and structural basis of these two systems. In light of the results presented in this paper, the thumb region and the larger substrate binding cleft of NpXyn11A seem to play a major role on the activity of this enzyme. Its lower thermal stability may instead be caused by the higher flexibility of certain regions located further from the active site. Regions such as the N-ter, the loops located in the fingers region, the palm loop, and the helix loop seem to be less stable than in the hyper-thermostable EvXyn11TS. By identifying molecular regions that are critical for the stability of these enzymes, this study allowed us to identify promising targets for engineering GH-11 xylanases. Eventually, we identify NpXyn11A as the ideal host for grafting the thermostabilizing traits of EvXyn11TS.



Author(s):  
Shiro Fujishiro ◽  
Harold L. Gegel

Ordered-alpha titanium alloys having a DO19 type structure have good potential for high temperature (600°C) applications, due to the thermal stability of the ordered phase and the inherent resistance to recrystallization of these alloys. Five different Ti-Al-Ga alloys consisting of equal atomic percents of aluminum and gallium solute additions up to the stoichiometric composition, Ti3(Al, Ga), were used to study the growth kinetics of the ordered phase and the nature of its interface.The alloys were homogenized in the beta region in a vacuum of about 5×10-7 torr, furnace cooled; reheated in air to 50°C below the alpha transus for hot working. The alloys were subsequently acid cleaned, annealed in vacuo, and cold rolled to about. 050 inch prior to additional homogenization



Author(s):  
W.F. Marshall ◽  
A.F. Dernburg ◽  
B. Harmon ◽  
J.W. Sedat

Interactions between chromatin and nuclear envelope (NE) have been implicated in chromatin condensation, gene regulation, nuclear reassembly, and organization of chromosomes within the nucleus. To further investigate the physiological role played by such interactions, it will be necessary to determine which loci specifically interact with the nuclear envelope. This will not only facilitate identification of the molecular determinants of this interaction, but will also allow manipulation of the pattern of chromatin-NE interactions to probe possible functions. We have developed a microscopic approach to detect and map chromatin-NE interactions inside intact cells.Fluorescence in situ hybridization (FISH) is used to localize specific chromosomal regions within the nucleus of Drosophila embryos and anti-lamin immunofluorescence is used to detect the nuclear envelope. Widefield deconvolution microscopy is then used to obtain a three-dimensional image of the sample (Fig. 1). The nuclear surface is represented by a surface-harmonic expansion (Fig 2). A statistical test for association of the FISH spot with the surface is then performed.



Author(s):  
Yih-Cheng Shih ◽  
E. L. Wilkie

Tungsten silicides (WSix) have been successfully used as the gate materials in self-aligned GaAs metal-semiconductor-field- effect transistors (MESFET). Thermal stability of the WSix/GaAs Schottky contact is of major concern since the n+ implanted source/drain regions must be annealed at high temperatures (∼ 800°C). WSi0.6 was considered the best composition to achieve good device performance due to its low stress and excellent thermal stability of the WSix/GaAs interface. The film adhesion and the uniformity in barrier heights and ideality factors of the WSi0.6 films have been improved by depositing a thin layer of pure W as the first layer on GaAs prior to WSi0.6 deposition. Recently WSi0.1 has been used successfully as the gate material in 1x10 μm GaAs FET's on the GaAs substrates which were sputter-cleaned prior to deposition. These GaAs FET's exhibited uniform threshold voltages across a 51 mm wafer with good film adhesion after annealing at 800°C for 10 min.



Author(s):  
J. Liu ◽  
N. D. Theodore ◽  
D. Adams ◽  
S. Russell ◽  
T. L. Alford ◽  
...  

Copper-based metallization has recently attracted extensive research because of its potential application in ultra-large-scale integration (ULSI) of semiconductor devices. The feasibility of copper metallization is, however, limited due to its thermal stability issues. In order to utilize copper in metallization systems diffusion barriers such as titanium nitride and other refractory materials, have been employed to enhance the thermal stability of copper. Titanium nitride layers can be formed by annealing Cu(Ti) alloy film evaporated on thermally grown SiO2 substrates in an ammonia ambient. We report here the microstructural evolution of Cu(Ti)/SiO2 layers during annealing in NH3 flowing ambient.The Cu(Ti) films used in this experiment were prepared by electron beam evaporation onto thermally grown SiO2 substrates. The nominal composition of the Cu(Ti) alloy was Cu73Ti27. Thermal treatments were conducted in NH3 flowing ambient for 30 minutes at temperatures ranging from 450°C to 650°C. Cross-section TEM specimens were prepared by the standard procedure.



2020 ◽  
Vol 477 (1) ◽  
pp. 173-189 ◽  
Author(s):  
Marco Pedretti ◽  
Carolina Conter ◽  
Paola Dominici ◽  
Alessandra Astegno

Arabidopsis centrin 2, also known as calmodulin-like protein 19 (CML19), is a member of the EF-hand superfamily of calcium (Ca2+)-binding proteins. In addition to the notion that CML19 interacts with the nucleotide excision repair protein RAD4, CML19 was suggested to be a component of the transcription export complex 2 (TREX-2) by interacting with SAC3B. However, the molecular determinants of this interaction have remained largely unknown. Herein, we identified a CML19-binding site within the C-terminus of SAC3B and characterized the binding properties of the corresponding 26-residue peptide (SAC3Bp), which exhibits the hydrophobic triad centrin-binding motif in a reversed orientation (I8W4W1). Using a combination of spectroscopic and calorimetric experiments, we shed light on the SAC3Bp–CML19 complex structure in solution. We demonstrated that the peptide interacts not only with Ca2+-saturated CML19, but also with apo-CML19 to form a protein–peptide complex with a 1 : 1 stoichiometry. Both interactions involve hydrophobic and electrostatic contributions and include the burial of Trp residues of SAC3Bp. However, the peptide likely assumes different conformations upon binding to apo-CML19 or Ca2+-CML19. Importantly, the peptide dramatically increases the affinity for Ca2+ of CML19, especially of the C-lobe, suggesting that in vivo the protein would be Ca2+-saturated and bound to SAC3B even at resting Ca2+-levels. Our results, providing direct evidence that Arabidopsis SAC3B is a CML19 target and proposing that CML19 can bind to SAC3B through its C-lobe independent of a Ca2+ stimulus, support a functional role for these proteins in TREX-2 complex and mRNA export.



2020 ◽  
Author(s):  
Feng Xiao ◽  
Bin Yao ◽  
Pavankumar Challa Sasi ◽  
Svetlana Golovko ◽  
Dana Soli ◽  
...  


1991 ◽  
Vol 1 (12) ◽  
pp. 1823-1836 ◽  
Author(s):  
M. Bessière ◽  
A. Quivy ◽  
S. Lefebvre ◽  
J. Devaud-Rzepski ◽  
Y. Calvayrac


1994 ◽  
Vol 4 (4) ◽  
pp. 653-657
Author(s):  
B. Bonzi ◽  
M. El Khomssi ◽  
H. Lanchon-Ducauquis


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