10 research outputs found

    Fungi hijack a ubiquitous plant apoplastic endoglucanase to release a ROS scavenging beta-glucan decasaccharide to subvert immune responses

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    Plant pathogenic and beneficial fungi have evolved several strategies to evade immunity and cope with host-derived hydrolytic enzymes and oxidative stress in the apoplast, the extracellular space of plant tissues. Fungal hyphae are surrounded by an inner insoluble cell wall layer and an outer soluble extracellular polysaccharide (EPS) matrix. Here, we show by proteomics and glycomics that these two layers have distinct protein and carbohydrate signatures, and hence likely have different biological functions. The barley (Hordeum vulgare) β-1,3-endoglucanase HvBGLUII, which belongs to the widely distributed apoplastic glycoside hydrolase 17 family (GH17), releases a conserved β-1,3;1,6-glucan decasaccharide (β-GD) from the EPS matrices of fungi with different lifestyles and taxonomic positions. This low molecular weight β-GD does not activate plant immunity, is resilient to further enzymatic hydrolysis by β-1,3-endoglucanases due to the presence of three β-1,6-linked glucose branches and can scavenge reactive oxygen species. Exogenous application of β-GD leads to enhanced fungal colonization in barley, confirming its role in the fungal counter-defensive strategy to subvert host immunity. Our data highlight the hitherto undescribed capacity of this often-overlooked EPS matrix from plant-associated fungi to act as an outer protective barrier important for fungal accommodation within the hostile environment at the apoplastic plant–microbe interface

    An Experimental Study of Mechanical Properties and Heat Transfer of Acrylic Composites with Structural and Surface Modified Al2O3 Particles

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    Thermal properties of acrylate-based composite combined with alumina-based particles were investigated. The composites were made of poly (methyl methacrylate) (PMMA) modified with dimethyl itaconate (DMI) as a matrix. Neat alumina particles (Al2O3) and alumina doped with iron oxide (Al2O3-Fe) modified with 3-aminopropyl-trimethoxylane (AM) and flax oil fatty acid methyl esters (biodiesel - BD) were used as reinforcement. Thermal conductivity measurements showed that composite with alumina particles with the highest alpha phase content had the highest thermal conductivity values. Mechanical properties (tensile strength, modulus of elasticity and elongation at break) were improved by the addition of modified alumina particles to PMMA/DMI matrix. The thermal properties are the most improved for composite with 3 wt.% Al2O3-Fe-AM particles that had the lowest temperature in a crack zone was noticed (10.28 %) compared to PMMA added due to the better stiffness and highest thermal conductivity

    N-Degradomic Analysis Reveals a Proteolytic Network Processing the Podocyte Cytoskeleton

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    Regulated intracellular proteostasis, controlled in part by proteolysis, is essential in maintaining the integrity of podocytes and the glomerular filtration barrier of the kidney. We applied a novel proteomics technology that enables proteome-wide identification, mapping, and quantification of protein N-termini to comprehensively characterize cleaved podocyte proteins in the glomerulus in vivo. We found evidence that defined proteolytic cleavage results in various proteoforms of important podocyte proteins, including those of podocin, nephrin, neph1, α-actinin-4, and vimentin. Quantitative mapping of N-termini demonstrated perturbation of protease action during podocyte injury in vitro, including diminished proteolysis of α-actinin-4. Differentially regulated protease substrates comprised cytoskeletal proteins as well as intermediate filaments. Determination of preferential protease motifs during podocyte damage indicated activation of caspase proteases and inhibition of arginine-specific proteases. Several proteolytic processes were clearly site-specific, were conserved across species, and could be confirmed by differential migration behavior of protein fragments in gel electrophoresis. Some of the proteolytic changes discovered in vitro also occurred in two in vivo models of podocyte damage (WT1 heterozygous knockout mice and puromycin aminonucleoside–treated rats). Thus, we provide direct and systems-level evidence that the slit diaphragm and podocyte cytoskeleton are regulated targets of proteolytic modification, which is altered upon podocyte damage
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