Soluble Protein Research Articles

Soluble form of the MDA5 protein in human sera

Viral double-stranded RNA (dsRNA) is sensed by toll-like receptor 3 (TLR3) and retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs), including melanoma differentiation-associated gene 5 (MDA5). MDA5 recognizes the genome of dsRNA viruses and replication intermediates of single-stranded RNA viruses. MDA5 also plays an important role in the development of autoimmune diseases, such as Aicardi-Goutieres syndrome and type I diabetes. Patients with dermatomyositis with serum MDA5 autoantibodies (anti-CADM-140) are known to have a high risk of developing rapidly progressive interstitial lung disease and poor prognosis. However, there have been no reports on the soluble form of MDA5 in human serum. In the present study, we generated in-house monoclonal antibodies (mAbs) against human MDA5. We then performed immunohistochemical analysis and sensitive sandwich immunoassays to detect the MDA5 protein using two different mAbs (clones H27 and H46). As per the immunohistochemical analysis, the MDA5 protein was moderately expressed in the alveolar epithelia of normal lungs and was strongly expressed in the cytoplasm of lymphoid cells in the tonsils and acinar cells of the pancreas. Interestingly, soluble MDA5 protein was detectable in the serum, but not in the urine, of healthy donors. Soluble MDA5 protein was also detectable in the serum of patients with dermatomyositis. Immunoblot analysis showed that human cells expressed a 120 kDa MDA5 protein, while the 60 kDa MDA5 protein increased in the supernatant of peripheral mononuclear cells within 15 min after MDA5 agonist/double-strand RNA stimulation. Hydrogen deuterium exchange mass spectrometry revealed that an anti-MDA5 mAb (clone H46) bound to the epitope (415QILENSLLNL424) derived from the helicase domain of MDA5. These results indicate that a soluble MDA5 protein containing the helicase domain of MDA5 could be rapidly released from the cytoplasm of tissues after RNA stimulation.

CK1δ/ε kinases regulate TDP-43 phosphorylation and are therapeutic targets for ALS-related TDP-43 hyperphosphorylation

Hyperphosphorylated TAR DNA-binding protein 43 (TDP-43) aggregates in the cytoplasm of neurons is the neuropathological hallmark of amyotrophic lateral sclerosis (ALS) and a group of neurodegenerative diseases collectively referred to as TDP-43 proteinopathies that includes frontotemporal dementia, Alzheimer's disease, and limbic onset age-related TDP-43 encephalopathy. The mechanism of TDP-43 phosphorylation is poorly understood. Previously we reported casein kinase 1 epsilon gene (CSNK1E gene encoding CK1ε protein) as being tightly correlated with phosphorylated TDP-43 (pTDP-43) pathology. Here we pursued studies to investigate in cellular models and in vitro how CK1ε and CK1δ (a closely related family sub-member) mediate TDP-43 phosphorylation in disease. We first validated the binding interaction between TDP-43 and either CK1δ and CK1ε using kinase activity assays and predictive bioinformatic database. We utilized novel inducible cellular models that generated translocated phosphorylated TDP-43 (pTDP-43) and cytoplasmic aggregation. Reducing CK1 kinase activity with siRNA or small molecule chemical inhibitors resulted in significant reduction of pTDP-43, in both soluble and insoluble protein fractions. We also established CK1δ and CK1ε are the primary kinases that phosphorylate TDP-43 compared to CK2α, CDC7, ERK1/2, p38α/MAPK14, and TTBK1, other identified kinases that have been implicated in TDP-43 phosphorylation. Throughout our studies, we were careful to examine both the soluble and insoluble TDP-43 protein fractions, the critical protein fractions related to protein aggregation diseases. These results identify CK1s as critical kinases involved in TDP-43 hyperphosphorylation and aggregation in cellular models and in vitro, and in turn are potential therapeutic targets by way of CK1δ/ε inhibitors.

Physiology and transcriptome analysis of Artemisia argyi adaptation and accumulation to soil cadmium

The soil pollution caused by cadmium (Cd) poses a significant threat to the environment. Therefore, identifying plants that can effectively remediate Cd-contaminated soils is urgently needed. In this study, physiological, cytological, and transcriptome analyses were performed to comprehensively understand the changes in Artemisia argyi under Cd stress. Physiological and cytological analyses indicated that A. argyi maintained normal growth with intact cell structure under Cd stress levels up to 10 mg/kg. Cytological analysis showed that Cd precipitation in leaf cells occurred in the cytoplasm and intercellular spaces. As the levels of Cd stress increased, proline accumulation in leaves increased, whereas soluble protein and soluble sugar initially increased, followed by a subsequent decline. The translocation factor was above 1 under 0.6 mg/kg Cd stress but decreased when it exceeded this concentration. Transcriptome analyses revealed several crucial Cd-influenced pathways, including amino acid, terpenoid, flavonoid, and sugar metabolisms. This study not only proved that A. argyi could enrich Cd in soil but also revealed the response of A. argyi to Cd and its resistance mechanisms, which provided insight into the cleaner production of A. argyi and the remediation of Cd-contaminated soil.

Dynamics of industrial hemp vegetative growth and metabolite accumulation in response to light intensity

Industrial hemp (Cannabis sativa L.) is a high-yielding cash crop with strong stress resistance, and its fibrous and metabolic profile holds significant development potential. In this study, we measured various agronomic traits, amino acids, saccharides, and their derivatives under four different light intensities (30 μmol·m−2·s−1, 80 μmol·m−2·s−1, 130 μmol·m−2·s−1, 180 μmol·m−2·s−1). Results showed that plant height decreased with increasing light intensity and reached a minimum at 180 μmol·m−2·s−1, which was 27 % less than the control treatment. Further, stem diameter increased by 52 %, leaf fresh weight by 130 %, stem fresh weight by 116 %, and root fresh weight by 264 % were at their maximum at 130 μmol·m−2·s−1. Plant dry weight of leaf (113 %), stem (214 %), and root (427 %) were also found to be maximum at 130 μmol·m−2·s−1. Metabolic results revealed that the free amino acid content was 10 %, 61 %, and 60 % higher under 30 μmol·m−2·s−1 compared to 80, 130, and 180 μmol·m−2·s−1. Additionally, soluble protein contents were 42 %, 81 %, and 77 % higher under 30 μmol·m−2·s−1 compared to 80, 130, and 180 μmol·m−2·s−1. In contrast, the soluble sugar contents were 42 %, 61 %, and 75 % higher than in the other three treatments. In conclusion, industrial hemp morphological characteristics and metabolite profiles exhibit distinct responses to different light intensities. To achieve desired outcomes, it is imperative to carefully choose an optimized light intensity specifically tailored to align with the defined objectives. These findings provide a foundation for a deeper understanding of the characteristics of industrial hemp, optimizing cultivation techniques, and investigating the impact of light intensity on industrial hemp vegetative period growth, amino acids, and saccharides.

Fractionation and characterisation of sialylated-mucin glycoprotein from edible birds' nest hydrolysates through anion exchange chromatography

Edible bird's nest (EBN) is made up of sialylated-mucin glycoprotein with various health benefits due to its high antioxidative activity. However, as a macromolecule with distinct charged sialic acid and amino acids, fractions with different charges would have varied physicochemical properties and antioxidant activity, which have not been studied. Therefore, this study aimed to fractionate and purify the enzymatic hydrolysed of cleaned EBN (EBNhc) and EBN by-product (EBNhbyp) through anion exchange chromatography (AEC), and determine their molecular weights, physicochemical properties, and antioxidative activities. Overall, 26 fractionates were collected from enzymatic hydrolysate by AEC, which were classified into 5 fractions. It was found that the positively charged fraction of EBNhc (CF1) and EBNhbyp (DF1) showed the significantly highest (p < 0.05) soluble protein contents (22.86 and 18.40 mg/g), total peptide contents (511.13 and 800.47 mg/g) and ferric reducing antioxidant power (17.44 and 6.96 mg/g) among the fractionates. In conclusion, a positively charged fraction (CF1 and DF 1) showed more desired physicochemical properties and antioxidative activities. This research suggests the potential of AEC fractionation as a technology to purify EBN and produce positively charged EBN fractionates with antioxidative potential that could be applied as food components to provide health benefits.

Engineering G protein-coupled receptors for stabilization.

G protein-coupled receptors (GPCRs) are one of the most important families of targets for drug discovery. One of the limiting steps in the study of GPCRs has been their stability, with significant and time-consuming protein engineering often used to stabilize GPCRs for structural characterization and drug screening. Unfortunately, computational methods developed using globular soluble proteins have translated poorly to the rational engineering of GPCRs. To fill this gap, we propose GPCR-tm, a novel and personalized structurally driven web-based machine learning tool to study the impacts of mutations on GPCR stability. We show that GPCR-tm performs as well as or better than alternative methods, and that it can accurately rank the stability changes of a wide range of mutations occurring in various types of class A GPCRs. GPCR-tm achieved Pearson's correlation coefficients of 0.74 and 0.46 on 10-fold cross-validation and blind test sets, respectively. We observed that the (structural) graph-based signatures were the most important set of features for predicting destabilizing mutations, which points out that these signatures properly describe the changes in the environment where the mutations occur. More specifically, GPCR-tm was able to accurately rank mutations based on their effect on protein stability, guiding their rational stabilization. GPCR-tm is available through a user-friendly web server at https://biosig.lab.uq.edu.au/gpcr_tm/.

A synergistic catalytic system of keratinases and disulfide reductases for the bioconversion of feather wastes to feed alternative protein

Effective hydrolysis of feather waste by keratinase is an important way to develop high-quality alternative proteins for feed. It has been shown that the reduction of disulfide bonds is the key rate-limiting step in the hydrolysis of feathers by keratinases. However, the reductive activity of keratinase itself is low. In order to increase the reductive activity of the catalytic system, we screened endogenous disulfide reductase from Bacillus subtilis and constructed a synergistic catalytic system, KerZ1/yjbI-S2-P1, consisting of keratinase and disulfide reductase by co-expression optimization. The reductive activity and feather hydrolytic activity of the synergistic system were 5.67-fold and 5-fold higher than the original system. Feathers were hydrolyzed to a higher extent compared with the original system. The total amino acid and soluble protein concentrations in the products increased by 99% and 59%, respectively. This study provides a more environmentally friendly and efficient reduction capability for keratinase to hydrolysis waste feathers, and also demonstrates a new way of alternative protein feedstock mining.

Coating seeds with biopolymers extracted from waste-activated sludge

Seeds can be coated with biopolymers extracted from waste-activated sludge, and the seeds of lettuce and spinach can be coated. When coated with the biopolymers, these seeds exhibited largely improved resistance to high temperature and humidity. The seeds coated or not coated with the biopolymers were aged, and the germination potential, germination ratio, and antioxidant enzyme activity of the coated seeds exhibited markedly greater differences than did those of the uncoated seeds. Moreover, the CAT, POD, and APX activities in the coated lettuce and spinach seeds increased by 56%, 2%, and 17%, respectively (lettuce), as did the activity of 8%, 3%, and 31%, respectively (spinach). In the pot trials, the contents of soluble protein and soluble sugar and the activities of antioxidant enzymes were also enhanced—42% and 56% (lettuce) as well as 34% and 23% (spinach). Thus, biopolymers can contribute to protecting against the aging of some rare seeds.

Elevated Temperature Effects on Protein Turnover Dynamics in Arabidopsis thaliana Seedlings Revealed by 15N-Stable Isotope Labeling and ProteinTurnover Algorithm

Global warming poses a threat to plant survival, impacting growth and agricultural yield. Protein turnover, a critical regulatory mechanism balancing protein synthesis and degradation, is crucial for the cellular response to environmental changes. We investigated the effects of elevated temperature on proteome dynamics in Arabidopsis thaliana seedlings using 15N-stable isotope labeling and ultra-performance liquid chromatography-high resolution mass spectrometry, coupled with the ProteinTurnover algorithm. Analyzing different cellular fractions from plants grown under 22 °C and 30 °C growth conditions, we found significant changes in the turnover rates of 571 proteins, with a median 1.4-fold increase, indicating accelerated protein dynamics under thermal stress. Notably, soluble root fraction proteins exhibited smaller turnover changes, suggesting tissue-specific adaptations. Significant turnover alterations occurred with redox signaling, stress response, protein folding, secondary metabolism, and photorespiration, indicating complex responses enhancing plant thermal resilience. Conversely, proteins involved in carbohydrate metabolism and mitochondrial ATP synthesis showed minimal changes, highlighting their stability. This analysis highlights the intricate balance between proteome stability and adaptability, advancing our understanding of plant responses to heat stress and supporting the development of improved thermotolerant crops.

A comparative biochemical characterization of Allium genetic resources of Türkiye

AbstractThe present study aimed to determine the variation in onion and garlic genetic resources collected from the Kelkit Valley, a unique biodiversity concerved region in Türkiye. For this purpose, 17 onion genotypes and 12 garlic genotypes were collected from local growers. A total of 9 biochemical analyses (total phenolic matter, total soluble protein, total dry matter, total flavonoid matter, ferric reducing antioxidant power, cupric reducing antioxidant capacity, 2,2-diphenyl-1-picrylhydrazylradical scavenging activity, hydrogen peroxide scavenging capacity and ascorbic acid) were performed on bulb and leaf extracts of all genotypes. According to variance analysis of the biochemical results from the samples, the differences between genotypes for all parameters were found to be significant (p &lt; 0.05). A post-hoc test (Tukey) and multiple means comparison were performed to analyze the results between groups in cases of significant differences. The results indicated that there is a large variation between different genotypes of both species and between both species and organs.

Protecting Lyophilized Escherichia coli Adenylate Kinase.

Drying protein-based drugs, usually via lyophilization, can facilitate storage at ambient temperature and improve accessibility but many proteins cannot withstand drying and must be formulated with protective additives called excipients. However, mechanisms of protection are poorly understood, precluding rational formulation design. To better understand dry proteins and their protection, we examine Escherichia coli adenylate kinase (AdK) lyophilized alone and with the additives trehalose, maltose, bovine serum albumin, cytosolic abundant heat soluble protein D, histidine, and arginine. We apply liquid-observed vapor exchange NMR to interrogate the residue-level structure in the presence and absence of additives. We pair these observations with differential scanning calorimetry data of lyophilized samples and AdK activity assays with and without heating. We show that the amino acids do not preserve the native structure as well as sugars or proteins and that after heating the most stable additives protect activity best.

Phenotype, Biomass, Carbon and Nitrogen Assimilation, and Antioxidant Response of Rapeseed under Salt Stress

Salt stress is one of the major adverse factors affecting plant growth and crop production. Rapeseed is an important oil crop, providing high-quality edible oil for human consumption. This experiment was conducted to investigate the effects of salt stress on the phenotypic traits and physiological processes of rapeseed. The soil salinity was manipulated by setting three different levels: 0 g NaCl kg−1 soil (referred to as S0), 1.5 g NaCl kg−1 soil (referred to as S1), and 3.0 g NaCl kg−1 soil (referred to as S2). In general, the results indicated that the plant height, leaf area, and root neck diameter decreased with an increase in soil salinity. In addition, the biomass of various organs at all growth stages decreased as soil salinity increased from S0 to S2. The increasing soil salinity improved the distribution of biomass in the root and leaf at the seedling and flowering stages, indicating that rapeseed plants subjected to salt stress during the vegetative stage are capable of adapting their growth pattern to sustain their capacity for nutrient and water uptake, as well as leaf photosynthesis. However, as the soil salinity increased, there was a decrease in the distribution of biomass in the pod and seed at the maturity stage, while an increase was observed in the root and stem, suggesting that salt stress inhibited carbohydrate transport into reproductive organs. Moreover, the C and N accumulation at the flowering and maturity stages exhibited a reduction in direct correlation with the increase in soil salinity. High soil salinity resulted in a reduction in the C/N, indicating that salt stress exerted a greater adverse effect on C assimilation compared to N assimilation, leading to an increase in seed protein content and a decrease in oil content. Furthermore, as soil salinity increased from S0 to S2, the activity of superoxide dismutase (SOD) and catalase (CAT) and the content of soluble protein and sugar increased by 58.39%, 33.38%, 15.57%, and 13.88% at the seedling stage, and 38.69%, 22.85%, 12.04%, and 8.26% at the flowering stage, respectively. In summary, this study revealed that salt stress inhibited C and N assimilation, leading to a suppressed phenotype and biomass accumulation. The imbalanced C and N assimilation under salt stress contributed to the alterations in the seed oil and protein content. Rapeseed had a certain degree of salt tolerance by improving antioxidants and osmolytes.

Characterization and application of recombinant Bovine Leukemia Virus Env protein

The Bovine Leukemia Virus (BLV) Envelope (Env) glycoprotein complex is instrumental in viral infectivity and shapes the host’s immune response. This study presents the production and characterization of a soluble furin-mutated BLV Env ectodomain (sBLV-EnvFm) expressed in a stable S2 insect cell line. We purified a 63 kDa soluble protein, corresponding to the monomeric sBLV-EnvFm, which predominantly presented oligomannose and paucimannose N-glycans, with a high content of core fucose structures. Our results demonstrate that our recombinant protein can be recognized from specific antibodies in BLV infected cattle, suggesting its potential as a powerful diagnostic tool. Moreover, the robust humoral immune response it elicited in mice shows its potential contribution to the development of subunit-based vaccines against BLV.

AggreProt: a web server for predicting and engineering aggregation prone regions in proteins.

Recombinant proteins play pivotal roles in numerous applications including industrial biocatalysts or therapeutics. Despite the recent progress in computational protein structure prediction, protein solubility and reduced aggregation propensity remain challenging attributes to design. Identification of aggregation-prone regions is essential for understanding misfolding diseases or designing efficient protein-based technologies, and as such has a great socio-economic impact. Here, we introduce AggreProt, a user-friendly webserver that automatically exploits an ensemble of deep neural networks to predict aggregation-prone regions (APRs) in protein sequences. Trained on experimentally evaluated hexapeptides, AggreProt compares to or outperforms state-of-the-art algorithms on two independent benchmark datasets. The server provides per-residue aggregation profiles along with information on solvent accessibility and transmembrane propensity within an intuitive interface with interactive sequence and structure viewers for comprehensive analysis. We demonstrate AggreProt efficacy in predicting differential aggregation behaviours in proteins on several use cases, which emphasize its potential for guiding protein engineering strategies towards decreased aggregation propensity and improved solubility. The webserver is freely available and accessible at https://loschmidt.chemi.muni.cz/aggreprot/.

Screening Carbon Nano Materials for Preventing Amyloid Protein Aggregation by Adopting a Facile Method.

The soluble-to-toxic transformation of intrinsically disordered amyloidogenic proteins such as amyloid beta (Aβ), α-synuclein, mutant Huntingtin Protein (mHTT) and islet amyloid polypeptide (IAPP) among others are associated with disorders such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD) and Type 2 Diabetes (T2D), respectively. The dissolution of mature fibrils and toxic amyloidogenic intermediates, including oligomers, continues to be the pinnacle in the treatment of neurodegenerative disorders. Yet, methods to effectively and quantitatively report on the interconversion between amyloid monomers, oligomers and mature fibrils fall short. Here we describe a simplified method that implements the use of gel electrophoresis to address the transformation between soluble monomeric amyloid proteins and mature amyloid fibrils. The technique implements an optimized but well-known, simple, inexpensive, and quantitative assessment previously used to assess the oligomerization of amyloid monomers and subsequent amyloid fibrils. This method facilitates the screening of small molecules that disintegrate oligomers and fibrils into monomers, dimers, and trimers and/or retain amyloid proteins in their monomeric forms. Most importantly, our optimized method diminishes existing barriers associated with existing (alternative) techniques to evaluate fibril formation and intervention.

Large-Area Silicon Nitride Nanosieve for Enhanced Diffusion-Based Exosome Isolation.

Nanoporous membranes have a variety of applications, one of which is the size-selective separation of nanoparticles. In drug delivery, nanoporous membranes are becoming increasingly important for the isolation of exosomes, which are bio-nanoparticles. However, the low pore density and thickness of commercial membranes limit their efficiency. There have been many attempts to fabricate sub-micrometer thin membranes, but the limited surface area has restricted their practicality. In this study, large-area silicon nitride nanosieves for enhanced diffusion-based isolation of exosomes are presented. Notably, these nanosieves are scaled to sizes of up to 4-inch-wafers, a significant achievement in overcoming the fabrication challenges associated with such expansive areas. The method employs a 200nm porous sieve (38.2% porosity) for exosome separation and a 50nm sieve (10.7% porosity) for soluble protein removal. These 300nmthick nanosieves outperform conventional polycarbonate membranes by being 50 times thinner, thereby increasing nanoparticle permeability. The method enables a 90% recovery rate of intact exosomes from human serum and a purity ratio of 3 × 107 particles/µg protein, 4.6 times higher than ultracentrifugation methods. The throughput of the method is up to 15mL by increasing the size of the nanosieve, making it an ideal solution for large-scale exosome production for therapeutic purposes.

Gelatinization and fermentation synergy: investigating the protein digestibility, mineral bioaccessibility and microstructural transformations of black mash beans through Saccharomyces cerevisiae and Lactobacillus spp

ABSTRACT This study explored the nutritional qualities of black mash bean flour impacted by fermentation, gelatinization and their combination. Protease activity, in vitro protein digestibility (IVPD), and protein solubility were evaluated for both fermented and unfermented black mash bean flour samples. Interestingly, when compared to fermented samples and gelatinized mash bean flour (GMBF), raw mash bean flour (RMBF) showed the highest protein solubility. Due to enzymatic activity and protein hydrolysis, fermentation with Lactobacillus E14 and Saccharomyces cerevisiae MK-157 enhanced protein solubility. Prolonged fermentation timeframes were positively correlated with protease activity and IVPD, suggesting the degradation of complex proteins. The highest levels of IVPD (90.12%) and protease activity (5546.78 U/g) were detected in GMBF fermented by S. cerevisiae MK-157, indicating the impact of fermentation on protein breakdown. The Lactobacillus E14-fermented RMBF exhibited 5521.45 U/g of protease activity and 87.54% IVPD. The mineral content of the fermented samples was substantially greater than that of the unfermented GMBF. The increased calcium, magnesium, potassium, sodium, iron, zinc, and copper levels for the GMBF fermented by S. cerevisiae MK-157 indicated that fermentation facilitated the release of minerals from chelated compounds. Microscopic examination revealed higher protein quantities in the fermented flour as well as hydrolyzed and broken starch granules embedded in the protein matrix. Large mash bean proteins were shown to be proteolyzed during fermentation into smaller peptides by SDS PAGE. Storage proteins were extensively broken down during fermentation with S. cerevisiae MK-157, whereas Lactobacillus E14 displayed a more focused pattern of hydrolysis. The nutritional profile of black mash bean flour was improved by fermentation with Lactobacillus E14 and S. cerevisiae MK-157.

Effect of ultrasound-assisted L-lysine treatment on pork meat quality and myofibrillar protein properties during postmortem aging.

This paper aimed to investigate the effects of ultrasound-assisted L-lysine treatment on meat quality and myofibrillar proteins (MPs) properties of pork longissimus dorsi during postmortem aging. The results revealed that the L-lysine (Lys) and/or ultrasound treatment significantly increased (p < 0.05) the water-holding capacity and tenderness of the pork during postmortem aging, while the ultrasound-assisted Lys treatment had the lowest cooking loss, pressurization loss, Warner-Bratzler shear force, and hardness. In addition, L-lysine and/or ultrasound treatment increased (p < 0.05) pH value, T21, and myofibrillar fragmentation index, while the ultrasound-assisted Lys treatment had the highest value. Meanwhile, the protein solubility was increased with Lys and/or ultrasound treatment during postmortem aging, and ultrasound-assisted Lys treatment had the highest solubility, reaching 88.19%, 92.98%, and 91.73% at 0, 1, and 3 days, respectively. The result of protein conformational characteristics showed that Lys and/or ultrasound treatment caused the unfolding of the α-helix structure, resulting in the exposure of more hydrophobic amino acids and buried sulfhydryl groups, ultimately enhancing MPs solubility. In summary, ultrasound-assisted Lys treatment altered the structure of MPs, resulting in the enhancement of the water-holding capacity and tenderness of the pork. PRACTICAL APPLICATION: This study showed that ultrasound-assisted L-lysine (Lys) treatment could enhance the water-holding capacity and tenderness of pork during postmortem aging. The results might provide a reference for the application of ultrasound-assisted Lys treatment on the improvement of pork meat quality. To facilitate practical applications in production, the development of medium and large-sized ultrasound equipment for conducting small-scale and pilot experiments is crucial for future research.

Tardigrade secretory proteins protect biological structures from desiccation

Tardigrades, microscopic animals that survive a broad range of environmental stresses, express a unique set of proteins termed tardigrade-specific intrinsically disordered proteins (TDPs). TDPs are often expressed at high levels in tardigrades upon desiccation, and appear to mediate stress adaptation. Here, we focus on the proteins belonging to the secreted family of tardigrade proteins termed secretory-abundant heat soluble (“SAHS”) proteins, and investigate their ability to protect diverse biological structures. Recombinantly expressed SAHS proteins prevent desiccated liposomes from fusion, and enhance desiccation tolerance of E. coli and Rhizobium tropici upon extracellular application. Molecular dynamics simulation and comparative structural analysis suggest a model by which SAHS proteins may undergo a structural transition upon desiccation, in which removal of water and solutes from a large internal cavity in SAHS proteins destabilizes the beta-sheet structure. These results highlight the potential application of SAHS proteins as stabilizing molecules for preservation of cells.

Potentialities of Technosol-isolated PGPB consortium in promoting plant growth in lettuce seedlings

Abstract Background and aims Reducing land degradation and safeguarding agricultural productions ensures the provision of ecosystem services and economic welfare, as highlighted by the 2030 Sustainable Development Goals. Among the promising solutions to tackle these issues, the study investigates the use of Technosol-isolated PGPB, as a novel approach for enhancing plant growth and the capability to cope with soil salinization. Methods Several bacteria have been isolated from a Technosol in Naples (Italy), selected to produce a consortium, based on their PGP features and tested on lettuce. The promotion of lettuce growth was evaluated both in soil (sterilized and not-sterilized Technosols) and in water agar media differing in NaCl concentrations, focusing on total polyphenols, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, soluble proteins, and sugars, which were evaluated on seedlings. Results Germination kinetics differed among treatments in both soil and artificial media setups. In the Technosol trial, the PGPB treated seeds were the quickest to emerge, showing a higher radical scavenging activity and lower soluble carbohydrate content. In the salinity trial, total soluble proteins were significantly higher in primed seeds at null saline cocentration. Conclusion Our results indicate that the PGPB application has a beneficial outcome on the Technosol but under saline stress produces an inhibitory or a costly effect on the biochemistry and germination kinetics. However, Technosol PGPBs employement may open new research scenarios on their potential application considering Nature-Based Solutions and/or plant growth in degraded environments.