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Cord-Landwehr, S.

Publications and source records attributed to Cord-Landwehr, S..

6 recordsLinked to original sources

Chitinase-3-like protein 1 decodes chitosan acetylation patterns into toll-like receptor 2 signaling through heparan sulfate

Chitinase-3-like protein 1 (CHI3L1), which is associated with a wide range of inflammatory diseases, lacks chitinase activity but retains the ability to bind chitin and chitosan. Chitin is a major component of fungal cell walls, whereas chitosan is used in biomedicine. In addition to chitosan, CHI3L1 has been proposed to interact with heparan sulfate (HS), a highly sulfated glycosaminoglycan on mammalian cell surfaces. Here, we investigated how interactions with chitosan and HS regulate the pro-inflammatory activity of CHI3L1. Mapping of the chitin-binding cleft revealed preferential binding of CHI3L1 to chitosans with a regular acetylation pattern that, together with CHI3L1, promoted toll-like receptor 2 signaling. We further identified a dominant HS-binding site that recognizes a distinct HS sulfation code containing a coherent motif of N- and 6-O-sulfations. Mutation of this HS-binding site or impaired HS biosynthesis prevented CHI3L1 accumulation at the cell surface and abolished CHI3L1-mediated cell activation. Together, our findings establish HS as a critical co-receptor for CHI3L1 and reveal a pro-inflammatory cross-talk between HS, CHI3L1, and chitosan that may contribute to host defense against fungal pathogens and responses to chitosan-based biomaterials. These findings identify HS- and chitosan-dependent CHI3L1 signaling as a potential target for modulating inflammatory responses.

Molecular Biology↗

Immunoengineered Chitosanase-Produced Chitosan Oligomers for Elevating Plant Resistance to Viral Infection

Chitooligomers can act as plant biostimulants or biopesticides, but todays chitosan-based agro-biologics often lack sufficient efficacy. This is due to a lack of scalable production processes for structurally well-controlled chitosans combined with a limited understanding of structure-function relationships. Chitosans differ in their degree of polymerization (DP), fraction and pattern of acetylation (FA and PA). While the influence of DP and FA on antimicrobial and phytostimulatory properties is at least partially known, this is not yet the case for PA. PA can be partially controlled by using enzymatic rather than acid hydrolysis for oligomer production. We have used recombinant chitinases and chitosanases to hydrolyse a well-characterised chitosan polymer, and purified oligomers with different DP. We have structurally characterised the products and tested their abilities to protect tobacco from viral disease. Chitinase products were dominated by GlcNAc units at their reducing and non-reducing ends, with GlcN units dominating their centers, and v.v. for chitosanase products. While the chitinase-derived hydrolysates were inactive, the chitosanase-derived oligomers possessed elicitor and priming activities and protected plants from disease, and their activity increased with increasing DP. Clearly, the Bacillus chitosanase used is well-suited to set up a scalable production process for chitosan oligomers with promising agro-biologic properties. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=95 SRC="FIGDIR/small/731087v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@b50b59org.highwire.dtl.DTLVardef@1fc35c3org.highwire.dtl.DTLVardef@12db91aorg.highwire.dtl.DTLVardef@d3732f_HPS_FORMAT_FIGEXP M_FIG C_FIG

plant biology↗

Structural and chemical properties of insects' chitin-containing extracellular matrices

Insects body barriers rely on specialized extracellular matrices that protect against harmful environmental influences. The outer barrier is the cuticle, which is composed of chitin, cuticle proteins and lipids. The peritrophic matrix (PM) serves as an inner barrier lining the midgut epithelium. It is composed of chitin fibers that are organized by PM proteins. While cuticle and PM proteins have received considerable attention in the past, supramolecular organization and physicochemical properties of the chitin component - particularly of the PM - remain poorly understood. Here, we combine synchrotron-based X-ray diffraction data from the PMs of lepidopteran and coleopteran insects with RNA interference (RNAi), mass spectrometric and histochemical analyses of the PM from Tribolium castaneum to determine chitins allomorphic state and degree of acetylation. The chitin of the PM exhibits signatures characteristic of dihydrate {beta}-chitin along the entire midgut. In contrast, the cuticle is made of tightly packed -chitin nanofibrils. Mass spectrometry revealed that the PMs chitin is highly acetylated (>95%). RNAi silencing of gut-specific genes encoding chitin deacetylasesTcCDA6-9 further increases the degree of acetylation. Histochemical analyses staining chitin with different degrees of acetylation confirm the predominance of highly acetylated chitin in the PM. Notably, the larval cuticle has a layered organization with deacetylated chitin present in exo- and highly acetylated chitin in endocuticles. Depletion of both TcCDA1 or TcCDA2 impairs chitin deacetylation, which indicates that both proteins cooperate in their activity in the integument. These results establish fundamental principles of polysaccharide-based extracellular matrices, with broad implications for insect biology.

molecular biology↗

Comparative chemical characterisation of chitosans and their impact on growth, faecal consistency and microbiota composition in weaned piglets

Managing post-weaning diarrhoea (PWD) in piglets is difficult due to limits on antibiotics and zinc. Chitosan is emerging as a potential feed additive. We analysed a chito-oligosaccharide hydrochloride (COS-HCl), a low molecular weight (LMW) chitosan, and a medium molecular weight (MMW) chitosan, and assessed their effects on growth, faecal consistency, microbiota, and potential interference with enterotoxigenic Escherichia coli (ETEC). The three chitosans were characterised using {superscript 1}H-NMR, SEC-RI-MS, and SEC-RI-MALLS. COS-HCl had an Mw of 0.824 kDa; LMW and MMW showed Mw ranges of 14.4 kDa (0.3-30 kDa) and 116 kDa (15-600 kDa). Degrees of acetylation were 9.5%, 6.5%, and 15%. Two 42-day field studies evaluated average daily gain (ADG), faecal consistency, and microbiota. In the first trial, COS-HCl at 0.025-0.1% did not significantly affect ADG (-33 to - 12 g/d). In the second, LMW and MMW at 0.01% did not significantly change ADG (-7 and +3 g/d). Faecal consistency, ETEC shedding, and microbiota composition were similar to controls. An enzymatic HPLC-MS method enabled quantification of MMW chitosan in premix. Our results highlight the importance of advanced chitosan characterisation for precision nutrition and suggest that a threshold dosemay be needed to benefit growth and gut health in PWD management. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/714014v1_ufig1.gif" ALT="Figure 1"> View larger version (31K): org.highwire.dtl.DTLVardef@ffce80org.highwire.dtl.DTLVardef@4e3603org.highwire.dtl.DTLVardef@bc0c73org.highwire.dtl.DTLVardef@1d55472_HPS_FORMAT_FIGEXP M_FIG C_FIG

pharmacology and toxicology↗

Time-resolved transcriptomics reveal a mechanism of host niche defense: beneficial root endophytes deploy a host-protective antimicrobial GH18-CBM5 chitinase

Associations between plants and beneficial root-endophytic fungi enhance plant performance by improving nutrient uptake, abiotic stress tolerance and disease resistance. To successfully colonize different host plants and defend their host niche against competing microbes, but also to cooperate with beneficial bacterial members of the microbiota, root endophytes such as Sebacinales secrete a multitude of tightly regulated effector-proteins and carbohydrate-active enzymes. However, the functions, specificity, and regulation of these proteins remain poorly understood. In this study, we employ time-resolved transcriptomics to analyse the gene expression profiles of two Sebacinales members interacting with organisms from different kingdoms of life. We identified crucial genes for plant colonization and intermicrobial competition, including a fungal GH18-CBM5 chitinase specifically upregulated in response to the phytopathogenic fungus Bipolaris sorokiniana. This chitinase protects the plant hosts against the pathogen, reducing fungal biomass and disease symptoms in barley and Arabidopsis thaliana. Our findings shed light on interaction partner specific gene expression in Sebacinales endophytes, with potential applications in enhancing plant health and resilience. Bullet pointsO_LIBoth Serendipita indica (Si) and Serendipita vermifera (Sv) show similar transcriptional responses to three host species and the phytopathogen Bipolaris sorokiniana (Bs), indicating common interaction principles between Sebacinales and plant hosts or fungi. C_LIO_LIThese shared mechanisms involve the activation of effector genes like small secreted proteins and carbohydrate-active enzymes. C_LIO_LICooperation with beneficial bacteria elicits only minimal transcriptomic alterations in Sebacinales compared to plants and Bs. C_LIO_LISebacinales respond to Bs by upregulating a specific GH18-CBM5 chitinase unique to Basidiomycota within the fungal kingdom, inhibiting Bs growth and reducing disease symptoms in Arabidopsis thaliana and barley. C_LI

microbiology↗

Chitosan perception in Arabidopsis requires the chitin receptor AtCERK1 suggesting an improved model for receptor structure and function

Chitin, a linear polymer of N-acetyl-D-glucosamine, and chitosans, fully or partially deacetylated derivatives of chitin, are known to elicit defense reactions in higher plants. We compared the ability of chitin and chitosan oligomers and polymers (chitin oligomers with degree of polymerization 3 to 8; chitosan oligomers with degree of acetylation 0% to 35% and degree of polymerization 3 to 15; chitosan polymers with degree of acetylation 1% to 60% and degree of polymerization ~1300) to elicit an oxidative burst indicative of induced defense reactions in A. thaliana seedlings. Fully deacetylated chitosans were not able to trigger a response; elicitor activity increased with increasing degree of acetylation of chitosan polymers. Partially acetylated chitosan oligomers required a minimum degree of polymerization of 6 and at least four N-acetyl groups to trigger a response. Invariably, elicitation of an oxidative burst required the presence of the chitin receptor AtCERK1. Our results as well as previously published studies on chitin and chitosan perception in plants are best explained by a new general model of LysM-containing receptor complexes where two partners form a long, but off-set chitin-binding groove and are, thus, dimerized by one chitin or chitosan molecule, sharing a central GlcNAc unit with which both LysM domains interact. To verify this model and to distinguish it from earlier models, we assayed elicitor and inhibitor activities of selected partially acetylated chitosan oligomers with fully defined structures. In contrast to the initial \"continuous groove\", the original \"sandwich\", or the current \"sliding mode\" models for the chitin/chitosan receptor, the here proposed \"slipped sandwich\" model - which builds on these earlier models and represents a consensus combination of these - is in agreement with all experimental observations.

biochemistry↗