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The calcium pump (sarco/endoplasmic reticulum Ca2+-ATPase, SERCA) plays a major role in calcium homeostasis in muscle cells by clearing cytosolic Ca2+ during muscle relaxation. Active Ca2+ transport by SERCA involves the structural transition from a low-Ca2+ affinity E2 state toward a high-Ca2+ affinity E1 state of the pump. This structural transition is accompanied by the countertransport of protons to stabilize the negative charge and maintain the structural integrity of the transport sites and partially compensate for the positive charges of the two Ca2+ ions passing through the membrane. X-ray crystallography studies have suggested that a hydrated pore located at the C-terminal domain of SERCA serves as a conduit for proton countertransport, but the existence and function of this pathway have not yet been fully characterized. We used atomistic simulations to demonstrate that in the protonated E2 state and the absence of initially bound water molecules, the C-terminal pore becomes hydrated in the nanosecond timescale. Hydration of the C-terminal pore is accompanied by the formation of water wires that connect the transport sites with the cytosol. Water wires are known as ubiquitous proton-transport devices in biological systems, thus supporting the notion that the C-terminal domain serves as a conduit for proton release. Additional simulations showed that the release of a single proton from the transport sites induces bending of transmembrane helix M5 and the interaction between residues Arg762 and Ser915. These structural changes create a physical barrier against full hydration of the pore and prevent the formation of hydrogen-bonded water wires once proton transport has occurred through this pore. Together, these findings support the notion that the C-terminal proton release pathway is a functional element of SERCA and also provide a mechanistic model for its operation in the catalytic cycle of the pump. 相似文献
3.
Laura Cuy-Chaparro Michel David Bohrquez Gabriela Arvalo-Pinzn Jeimmy Johana Castaeda-Ramírez Carlos Fernando Surez Laura Pabn Diego Ordez Gina Marcela Gallego-Lpez Carlos Esteban Surez Darwin Andrs Moreno-Prez Manuel Alfonso Patarroyo 《International journal of molecular sciences》2021,22(2)
Apical membrane antigen 1 is a microneme protein which plays an indispensable role during Apicomplexa parasite invasion. The detailed mechanism of AMA-1 molecular interaction with its receptor on bovine erythrocytes has not been completely defined in Babesia bovis. This study was focused on identifying the minimum B. bovis AMA-1-derived regions governing specific and high-affinity binding to its target cells. Different approaches were used for detecting ama-1 locus genetic variability and natural selection signatures. The binding properties of twelve highly conserved 20-residue-long peptides were evaluated using a sensitive and specific binding assay based on radio-iodination. B. bovis AMA-1 ectodomain structure was modelled and refined using molecular modelling software. NetMHCIIpan software was used for calculating B- and T-cell epitopes. The B. bovis ama-1 gene had regions under functional constraint, having the highest negative selective pressure intensity in the Domain I encoding region. Interestingly, B. bovis AMA-1-DI (100YMQKFDIPRNHGSGIYVDLG119 and 120GYESVGSKSYRMPVGKCPVV139) and DII (302CPMHPVRDAIFGKWSGGSCV321)-derived peptides had high specificity interaction with erythrocytes and bound to a chymotrypsin and neuraminidase-treatment sensitive receptor. DI-derived peptides appear to be exposed on the protein’s surface and contain predicted B- and T-cell epitopes. These findings provide data (for the first-time) concerning B. bovis AMA-1 functional subunits which are important for establishing receptor-ligand interactions which could be used in synthetic vaccine development. 相似文献
4.
Bouaziz Youssef Royer Eric Bresson Guillaume Dhome Michel 《Multimedia Tools and Applications》2022,81(16):22449-22480
Multimedia Tools and Applications - Changes in appearance present a tremendous problem for the visual localization of an autonomous vehicle in outdoor environments. Data association between the... 相似文献
5.
Jing Huang Nairveen Ali Elsie Quansah Shuxia Guo Michel Noutsias Tobias Meyer-Zedler Thomas Bocklitz Jürgen Popp Ute Neugebauer Anuradha Ramoji 《International journal of molecular sciences》2021,22(4)
In recent decades, vibrational spectroscopic methods such as Raman and FT-IR spectroscopy are widely applied to investigate plasma and serum samples. These methods are combined with drop coating deposition techniques to pre-concentrate the biomolecules in the dried droplet to improve the detected vibrational signal. However, most often encountered challenge is the inhomogeneous redistribution of biomolecules due to the coffee-ring effect. In this study, the variation in biomolecule distribution within the dried-sample droplet has been investigated using Raman and FT-IR spectroscopy and fluorescence lifetime imaging method. The plasma-sample from healthy donors were investigated to show the spectral differences between the inner and outer-ring region of the dried-sample droplet. Further, the preferred location of deposition of the most abundant protein albumin in the blood during the drying process of the plasma has been illustrated by using deuterated albumin. Subsequently, two patients with different cardiac-related diseases were investigated exemplarily to illustrate the variation in the pattern of plasma and serum biomolecule distribution during the drying process and its impact on patient-stratification. The study shows that a uniform sampling position of the droplet, both at the inner and the outer ring, is necessary for thorough clinical characterization of the patient’s plasma and serum sample using vibrational spectroscopy. 相似文献
6.
Alfonso Martín-Bernab Josep Tarrag-Celada Valrie Cunin Sylvie Michelland Roldn Corts Johann Poignant Cyril Boyault Walid Rachidi Sandrine Bourgoin-Voillard Marta Cascante Michel Seve 《International journal of molecular sciences》2021,22(7)
Growing evidence is showing that acetylation plays an essential role in cancer, but studies on the impact of KDAC inhibition (KDACi) on the metabolic profile are still in their infancy. Here, we analyzed, by using an iTRAQ-based quantitative proteomics approach, the changes in the proteome of KRAS-mutated non-small cell lung cancer (NSCLC) A549 cells in response to trichostatin-A (TSA) and nicotinamide (NAM) under normoxia and hypoxia. Part of this response was further validated by molecular and biochemical analyses and correlated with the proliferation rates, apoptotic cell death, and activation of ROS scavenging mechanisms in opposition to the ROS production. Despite the differences among the KDAC inhibitors, up-regulation of glycolysis, TCA cycle, oxidative phosphorylation and fatty acid synthesis emerged as a common metabolic response underlying KDACi. We also observed that some of the KDACi effects at metabolic levels are enhanced under hypoxia. Furthermore, we used a drug repositioning machine learning approach to list candidate metabolic therapeutic agents for KRAS mutated NSCLC. Together, these results allow us to better understand the metabolic regulations underlying KDACi in NSCLC, taking into account the microenvironment of tumors related to hypoxia, and bring new insights for the future rational design of new therapies. 相似文献
7.
Solenn Percelay Jean-Marie Billard Thomas Freret Annie Andrieux Michel Boulouard Valentine Bouet 《International journal of molecular sciences》2021,22(5)
For a better translation from treatment designs of schizophrenia to clinical efficiency, there is a crucial need to refine preclinical animal models. In order to consider the multifactorial nature of the disorder, a new mouse model associating three factors (genetic susceptibility—partial deletion of the MAP6 gene, early-life stress—maternal separation, and pharmacological treatment—chronic Δ-9-tetrahydrocannabinol during adolescence) has recently been described. While this model depicts a schizophrenia-like phenotype, the neurobiological correlates remain unknown. Synaptic transmission and functional plasticity of the CA1 hippocampal region of male and female 3-hit mice were therefore investigated using electrophysiological recordings on the hippocampus slice. While basal excitatory transmission remained unaffected, NMDA receptor (NMDAr)-mediated long-term potentiation (LTP) triggered by theta-burst (TBS) but not by high-frequency (HFS) stimulation was impaired in 3-hit mice. Isolated NMDAr activation was not affected or even increased in female 3-hit mice, revealing a sexual dimorphism. Considering that the regulation of LTP is more prone to inhibitory tone if triggered by TBS than by HFS, the weaker potentiation in 3-hit mice suggests a deficiency of intrinsic GABA regulatory mechanisms. Indeed, NMDAr activation was increased by GABAA receptor blockade in wild-type but not in 3-hit mice. This electrophysiological study highlights dysregulations of functional properties and plasticity in hippocampal networks of 3-hit mice, one of the mechanisms suspected to contribute to the pathophysiology of schizophrenia. It also shows differences between males and females, supporting the sexual dimorphism observed in the disorder. Combined with the previously reported study, the present data reinforce the face validity of the 3-hit model that will help to consider new therapeutic strategies for psychosis. 相似文献
8.
Elham Shafiei Gol Mari-Klara Stein Michel Avital 《The Journal of Strategic Information Systems》2019,28(2):175-195
Crowdwork, a new form of digitally mediated employment and part of the so-called gig economy, has the capacity to change the nature of work organization and to provide strategic value to workers, job providers, and intermediary platform owners. However, because crowdwork is temporary, large-scale, distributed, and mediated, its governance remains a challenge that often casts a shadow over its strategic value. The objective of this paper is to shed light on the making of value-adding crowdwork arrangements. Specifically, the paper explores crowdwork platform governance mechanisms and the relationships between these mechanisms and organizational value creation. Building on a comprehensive review of the extant literature on governance and crowdwork, we construct an overarching conceptual model that integrates control system and coordination system as two complementary mechanisms that drive crowdwork platform governance effectiveness and the consequent job provider benefits. Furthermore, the model accentuates the role of the degree of centralization and the degree of routinization as critical moderators in crowdwork platform governance. Overall, the paper highlights the potential of crowdwork to contribute not only to inclusion, fair wages and flexible work arrangements for workers but also to organizations’ value and competitive edge. 相似文献
9.
10.
We developed a simple solution mixing and molding process for the incorporation of graphene nano-flakes (GNFs) in polymer films. Poly(vinylidene fluoride-co-hexafluoropropylene) (PVDF-HFP) and poly(ethylene-co-methacrylic acid) (PEMAA) were used for preparation of the samples. The orientation and stacking of GNFs were confirmed using a scanning electron microscope. The thermal conductivity values for these composites were obtained using (1) laser flash method (commercially available) and (2) an optothermal Raman (OTR) technique (homemade device). The former measures the thermal diffusivity (α) and one needs to measure the density (ρ) and the heat capacity (Cp) of the composites in order to measure the in-plane thermal conductivity (κ = α.ρ.Cp), while the latter measures the in-plane thermal conductivity directly from the relation between the excitation power and the position of the Raman resonance. The data obtained from Raman spectroscopy were analyzed, assuming heat propagation in three and two dimensions. The Raman results obtained based on the two-dimensional model were very close to the results obtained using the laser flash method with less than 10% difference. The OTRspectroscopy was found to be a promising technique for measuring the in-plane thermal conductivity of carbon-based polymer composites. PVDF-HFP and PEMAA composite films with very high in-plane thermal conductivity (25 W m−1 K−1) were obtained through the incorporation of GNFs (20 wt % concentration). Considering a very low thermal conductivity of these polymers (<0.2 W m−1 K−1), this corresponds to a large enhancement of roughly 12 400%. © 2020 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48927. 相似文献