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The advent of multi-specific targeted protein degradation (TPD) therapies has made it possible to drug targets that have long been considered to be inaccessible. For this reason, the foremost TPD modalities - molecular glues and proteolysis targeting chimeras (PROTACs) -have been widely adopted and developed in therapeutic programs across the pharmaceutical and biotechnology industries. While there are many clear advantages to these two approaches, there are also blind spots. Specifically, PROTACs and molecular glues are inherently mechanistically analogous in that targets of both are degraded via the 26s proteasome; however, not all disease-relevant targets are suitable for ubiquitin proteasome system (UPS)-mediated degradation. The alternative mammalian protein degradation pathway, the autophagy-lysosome system (or ALS), is capable of degrading targets that elude the UPS such as long-lived proteins, insoluble protein aggregates, and even abnormal organelles. Emerging TPD strategies- such as ATTEC, AUTAC, and LYTAC- take advantage of the substrate diversity of the ALS to greatly expand the clinical utility of TPD. In this Perspective, we will discuss the array of current TPD modalities, with a focus on critical evaluation of these novel ALS-mediated degradation techniques.

Feather is the main waste of poultry industries and constitutes of 90% keratin. Wastes composed of keratin are insoluble and recalcitrant to degradation using conventional decomposing methods. Microbial degradation for keratinous wastes is a promising approach for being eco-friendly and economically. However, due to insolubility of keratinous waste, it has several challenges in upstream and downstream processes such as culture medium optimization, designing of bioreactor, bioreaction/flow type, bioreactor configurations considering mass and heat transfer limitations, rheology derived problems, monitoring of microbial activity, choosing of the right scale-up parameter and purification. In this study, the challenges for keratin degradation processes were discussed with the aim of opening new opportunities for keratinous waste treatments in industrial level.

Protein aggregation is commonly observed phenomenon occurring during production of recombinant proteins in genetically engineered organisms over-expressing foreign genes. Aggregation of such recombinant proteins often leads to the formation of insoluble protein clusters named inclusion bodies (IBs). Although usually considered as waste by-products of protein production, over the last decade, this general misconception has been questioned by various studies. Gradual understanding of the structural, molecular and physiological features has revealed promising potential of these so far undesirable cellular products. In the present review we summarize basic characteristics of IBs, their use in antimicrobial peptides production and in various biotechnological fields, including tissue engineering, drug delivery and biocatalysis.

Aim of the present study was to evaluate the feeding value of spent coffee waste in ruminants. Nutrient composition of spent coffee waste (SCW) revealed that it contains Crude protein, Ether extract, Crude fibre and Nitrogen free extract at 12.02%, 9.23%, 56.20% and 21.73% respectively. SCW had higher neutral detergent fibre, acid detergent fibre and Lignin content of 79.67, 68.54 and 34.08% respectively when compared with conventional feeds and also contained high amount of Acid detergent insoluble protein (67.54%). Total phenol content in SCW was 0.35%. In vitro gas production studies revealed that SCW has significantly (P

Prion diseases are characterized by the self-templated misfolding of the cellular prion protein (PrPC) into infectious aggregates (PrPSc). The detailed molecular basis of the misfolding and aggregation of PrPC remains incompletely understood. It is believed that the transient misfolding of PrPC into partially structured intermediates precedes the formation of insoluble protein aggregates and is a critical component of the prion misfolding pathway. A number of environmental factors have been shown to induce the destabilization of PrPC and promote its initial misfolding. Recently, oxidative stress and reactive oxygen species (ROS) have emerged as one possible mechanism by which the destabilization of PrPC can be induced under physiological conditions. Methionine residues are uniquely vulnerable to oxidation by ROS and the formation of methionine sulfoxides leads to the misfolding and subsequent aggregation of PrPC. Here, we provide a review of the evidence for the oxidation of methionine residues in PrPC and its potential role in the formation of pathogenic prion aggregates.

Background Acid-induced hydrolysis of proteins has been used to improve the solubility and functional properties of various proteins, and could be a promising tool to facilitate the use of currently underutilized insoluble microalgae protein-rich fractions in food applications. However, the results of a prior study showed an unusual resistance of an insoluble microalgae protein-rich fraction to acid hydrolysis at room temperature. Results In the present study, the insoluble protein-rich fraction extracted from microalgae Chlorella prothothecoides was treated with 0.5 mol L-1 hydrochloric acid at 25, 45, 65 or 85 °C for 0-4 h. The results showed that hydrolysis of the fraction at 85 °C for 4 h led to decreases in the amount of insoluble protein-rich aggregates and the formation of fragments with a lower molecular weight, as well as an increase in protein solubility by approximately 40%. Nevertheless, some aggregated insoluble protein-rich particles remained, even after hydrolysis at 85 °C for 4 h. Conclusion The higher temperature improved the efficiency of the acid hydrolysis of the insoluble protein fraction from microalgae Chlorella prothothecoides, which is highly acid-resistant. Overall, an erosion-based mechanism was suggested for the acid hydrolysis of insoluble microalgae protein fraction. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

El objetivo fue desarrollar modelos de calibración de espectroscopia de reflectancia en el infrarrojo cercano (NIRS) para estimar el contenido de las fracciones de fibra y de carbohidratos en pastos Urochloa sp. para la zona tropical de México. Ciento ochenta y nueve muestras de tres híbridos de Urochloa fueron cosechadas cada 35 días durante un año, las cuales fueron analizadas por química húmeda para las fracciones de fibra: Fibra Detergente Neutra (NDF), Proteína insoluble en Detergente Neutro (NDIP), Fibra Detergente Ácida (ADF), Proteína Insoluble en Detergente Ácido (ADIP) y Lignina (LIG), y se estimaron los carbohidratos totales (TC), carbohidratos fibrosos (FC), carbohidratos no fibrosos (NFC) y carbohidratos indigestibles (UC). Los modelos de calibración y de validación se evaluaron por los respectivos Coeficientes de Determinación (R2); Errores Estándar de Validación Cruzada (SECV); y la desviación residual de la predicción (RPD). Además, con un conjunto de muestras externas, se realizó una prueba de sensibilidad evaluada por el Cuadrado Medio del Error de Predicción (MSPE). Las calibraciones obtenidas presentaron un adecuado ajuste y eficiencia predictiva mediante espectrofotometría NIR para NDF (RPD 2.4) y TC (RPD 2.5) lo que demuestra confiabilidad y capacidad de predicción para estas fracciones. Sin embargo, LIG, FC, NFC y UC requieren más observaciones para acrecentar su confiabilidad.

The aftermath of TBI is associated with an acute stress response and the accumulation of insoluble protein aggregates. Even after the symptoms of TBI are resolved, insidious molecular processes continue to develop, which often ultimately result in the development of age-associated neurodegenerative disorders. The precise molecular cascades that drive unhealthy brain aging are still largely unknown. In this review, we discuss proteostatic dysfunction as a converging mechanism contributing to accelerated brain aging after TBI. We examine evidence from human tissue and in vivo animal models, spanning both the aging and injury contexts. We conclude that TBI has a sustained debilitating effect on the proteostatic machinery, which may contribute to the accelerated pathological and cognitive hallmarks of aging that are observed following injury.

This study aimed to evaluate the effect of different Clitoria fairchildiana tree shading levels on the Tanganyika grass (Megatyrsus maximus Jacq. cv. Tanganyika) chemical composition, in summer I (2011/2012), spring (2012) and summer II (2012/2013) seasons. For this purpose, an experiment was conducted in a completely randomized design with four treatments (shading levels) and five repetitions at Seropédica, state of Rio de Janeiro, Brazil. The following variables were evaluated: dry matter (DM), crude protein (CP), neutral detergent (NDF) and acid detergent (ADF) fiber, neutral (NDIP) and acid (ADIP) detergent insoluble protein, lignin (LIG) and mineral matter (MM) contents. Data were analyzed by PROC MIXED SAS®, with repeated measures in time, and treatment means compared by Tukey’s test (P

Autophagy is a ubiquitous mechanism for maintaining cellular homeostasis through the degradation of long-lived proteins, insoluble protein aggregates, and superfluous or damaged organelles. Dysfunctional autophagy is observed in a variety of human diseases. With advanced research into the role that autophagy plays in physiological and pathological conditions, targeting autophagy is becoming a novel tactic for disease management. Saponins are naturally occurring glycosides containing triterpenoids or steroidal sapogenins as aglycones, and some saponins are reported to modulate autophagy. Research suggests that saponins may have therapeutic and preventive efficacy against many autophagy-related diseases. Therefore, this review comprehensively summarizes and discusses the reported saponins that exhibit autophagy regulating activities. In addition, the relevant signaling pathways that the mechanisms involved in regulating autophagy and the targeted diseases were also discussed. By regulating autophagy and related pathways, saponins exhibit bioactivities against cancer, neurodegenerative diseases, atherosclerosis and other cardiac diseases, kidney diseases, liver diseases, acute pancreatitis, and osteoporosis. This review provides an overview of the autophagy-regulating activity of saponins, the underlying mechanisms and potential applications for managing various diseases.

Aging animals accumulate insoluble protein as a consequence of a decline of proteostatic maintenance with age. In Caenorhabditis elegans, for instance, levels of detergent-insoluble protein increase with age. In longer-lived strains of C. elegans, this accumulation occurs more slowly, implying a link to lifespan determination. We further explored this link, and found that detergent-insoluble protein accumulates more rapidly at higher temperatures, a condition where lifespan is short. We employed a C. elegans strain carrying a GFP transcriptional reporter under the control of a heat shock (hsp-16.2) promoter to investigate the dynamics of proteostatic failure in individual nematodes. We found that early, sporadic activation of hsp-16.2 was predictive of shorter remaining lifespan in individual nematodes. Exposure to rapamycin, resulting in reduced mTOR signaling, delayed spurious expression, extended lifespan, and delayed accumulation of insoluble protein, suggesting that targets downstream of the mTOR pathway regulate the accumulation of insoluble protein. We specifically explored ribosomal S6 kinase (rsks-1) as one such candidate and found that RNAi against rsks-1 also resulted in less age-dependent accumulation of insoluble protein and extended lifespan. Our results demonstrate that inhibition of protein translation via reduced mTOR signaling resulted in slower accumulation of insoluble protein, delayed proteostatic crisis and extended lifespan in C. elegans.

Background All cells accumulate insoluble protein aggregates throughout their lifespan. While many studies have characterized the canonical disease-associated protein aggregates, such as those associated with amyloid plaques, additional, undefined proteins aggregate in the brain and may be directly associated with disease and lifespan. Methods A proteomics approach was used to identify a large subset of insoluble proteins in the mild cognitively impaired (MCI) and Alzheimer’s disease (AD) human brain. Cortical samples from control, MCI, and AD patients were separated into detergent-soluble and detergent-insoluble fractions, and high-resolution LC/MS/MS technology was used to determine which proteins became more insoluble in the disease state. Bioinformatics analyses were used to determine if the alteration of protein aggregation between AD and control patients was associated with any specific biological process. Western blots were used to validate the proteomics data and to assess the levels of secondary protein modifications in MCI and AD. Results There was a stage-dependent increase in detergent-insoluble proteins, with more extreme changes occurring in the AD cohort. Glycolysis was the most significantly overrepresented gene ontology biological process associated with the alteration of protein aggregation between AD and control patients. It was further shown that many low molecular weight proteins that were enriched in the AD brain were also highly aggregated, migrating on SDS-PAGE far above their predicted molecular masses. Glucose-6-phosphate isomerase, ubiquitin carboxyl-terminal hydrolase isoenzyme L1 (UCHL1/PARK5), and the DNA damage repair enzyme KU70 were among the top insoluble proteins identified by proteomics and validated by Western blot to be increased in the insoluble fractions of both MCI and AD brain samples. Conclusions Diverse proteins became more detergent-insoluble in the brains of both MCI and AD patients compared to age-matched controls, suggesting that multiple proteins aggregate in these diseases, likely posing a direct toxic insult to neurons. Furthermore, detergent-insoluble proteins included those with important biological activities for critical cellular processes such as energetics, proteolysis, and DNA damage repair. Thus, reduced protein solubility likely promotes aggregation and limits functionality, reducing the efficiency of multiple aspects of cell physiology. Pharmaceutical interventions that increase autophagy may provide a useful therapeutic treatment to combat protein aggregation.

Two different pearling degrees of wheat kernels (lightly-pearled: 14.4% and heavily-pearled: 38.9%) and un-pearled kernels were treated with ozone and evaluated for flour compositions and properties. Ozonation did not change main compositions and damaged starch content of three kernels' flours. Flour brightness of all three kernels was improved after ozone treatment. Ozonation enhanced the dough strength of the flours from un-pearled and pearled kernels and the effect elevated with increasing pearling degree. Ozone treatment increased the peak viscosity of flour and the level of increase in heavily-pearled kernels was greater than un-pearled and lightly-pearled. Ozonation resulted in an increase in the insoluble protein polymer content of heavily-pearled kernels' flour, but only had a slight effect on un-pearled lightly-pearled kernels. After ozone treatment, un-pearled and lightly-pearled kernels exhibited increases in molecular weight of starch, but heavily-pearled resulted in the opposite trend.

© 2020 by the authors., Age-dependent alterations in the proteostasis network are crucial in the progress of prevalent neurodegenerative diseases, such as Alzheimer’s, Parkinson’s, or amyotrophic lateral sclerosis, which are characterized by the presence of insoluble protein deposits in degenerating neurons. Because molecular chaperones deter misfolded protein aggregation, regulate functional phase separation, and even dissolve noxious aggregates, they are considered major sentinels impeding the molecular processes that lead to cell damage in the course of these diseases. Indeed, members of the chaperome, such as molecular chaperones and co-chaperones, are increasingly recognized as therapeutic targets for the development of treatments against degenerative proteinopathies. Chaperones must recognize diverse toxic clients of different orders (soluble proteins, biomolecular condensates, organized protein aggregates). It is therefore critical to understand the basis of the selective chaperone recognition to discern the mechanisms of action of chaperones in protein conformational diseases. This review aimed to define the selective interplay between chaperones and toxic client proteins and the basis for the protective role of these interactions. The presence and availability of chaperone recognition motifs in soluble proteins and in insoluble aggregates, both functional and pathogenic, are discussed. Finally, the formation of aberrant (pro-toxic) chaperone complexes will also be disclosed., This research was funded by the BBVA Foundation (grant number BBM-TRA-0203) and the Spanish AEI/EU-FEDER (grant number PID2019-109276RA-I00). J.O. is a Ramón y Cajal fellow from the Spanish AEI—Ministry of Science and Innovation.

Cytochrome P450 (CYP450) forms an enzyme superfamily that is ubiquitously distributed in nature and represents one of the largest and oldest gene superfamilies. CYP450 is important for the biotransformation of sterols and xenobiotics including drugs and is useful in the synthesis and degradation of signaling molecules. cyp450 gene encodes heme monooxygenases with specific substrate range and have numerous paralogues in most eukaryotes including free-living flatworms. This research work focuses on the human liver fluke Opisthorchis felineus, which is an emerging cause of hepatobiliary diseases such as cholangitis, obstructive jaundice, hepatomegaly, cholecystitis and cholelithiasis. The cyp450 gene was cloned in pET28a and was expressed in Escherichia coli. The insoluble of CYP450 was purified using a nickel affinity column with chelating Ni-NTA Sepharose matrix after solubilizing with sodium lauroyl sarcosinate (Sarkosyl). The affinity purified recombinant of CYP450 was further purified using size exclusion chromatography (SEC) to apparent homogeneity (~5 mg/L). Following SEC, of CYP450 was stable and observed to be a ~50kDa monomeric protein. Large quantities of the protein will guide in its biochemical and biophysical characterization, which may lead to the identification of new inhibitors of of CYP450.

In the present work the effect of the presence of soluble and insoluble protein on the stability of oil-water emulsions prepared with amaranth protein isolates (API) was analyzed. For this purpose, four types of emulsions were prepared: API-pH2 and API-pH6.3 dispersions and solutions. At pH 2.0 the amaranth proteins present higher solubility, are denatured and partially hydrolyzed; while at pH 6.3 its solubility is lower and its structure is more similar to the native protein. The soluble proteins present in API-pH2 and API-pH6.3 reduce with equal intensity the interfacial tension. However, the proteins present in API-pH2 are adsorbed twice as fast as those present in API-pH6.3, with equal rearrangement rate at the oil/water interface. Both, solutions and dispersions of API-pH2 and API-pH6.3 allow the formation of oil-in-water emulsions. Flocculation phenomena are evident, particularly in the case of API-pH6.3 dispersions. The calculated creaming-flocculation constant demonstrates that stability of emulsions increase with protein concentration and with the decrease of pH. The behaviour of API-pH2 and API-pH6.3 solutions was similar to that corresponding to the dispersions discarding a negative effect of the insoluble protein on the emulsifying properties of amaranth proteins., Centro de Investigación y Desarrollo en Criotecnología de Alimentos

A rapid shear-based test using a GlutoPeak instrument was compared with tests commonly used by durum wheat breeders to assess the potential of this instrument to discriminate between samples. Thirty-two durum wheat semolina samples were analyzed by mixograph, SDS sedimentation (SDSS), gluten index (GI), and GlutoPeak testing. A subset was also tested for pasta quality. GlutoPeak peak maximum time (PMT) was the best indicator of gluten strength and correlated well with the other tests except SDSS. Samples with higher levels of SDS-unextractable glutenin (insoluble protein [IP]) had stronger dough and longer PMT, but the GlutoPeak test only correlated with pasta stickiness using a smaller set of samples. The range in mixogram profiles encountered in breeding material was related to the IP content, and the pasta made from the different types was of similar quality, differing more because of protein content rather than mixogram type. The GlutoPeak test is faster than GI and uses less sample, requires little t...

The goal was to determine the ideal cutting age of elephant grass BRS Canará for forage production in the dry and rainy seasons. The experimental design comprised randomized blocks, with five treatments (42, 56, 70, 84, and 98 days in the dry season and 42, 60, 76, 91, and 105 days of regrowth in the rainy season) and four replications each. Agronomic, morphological, and chemical characteristics were evaluated. In the dry season, there was a significant quadratic response for plant height and leaf blade length. For the other characteristics, a significant linear response was observed, except stem diameter. In the rainy season, a quadratic response for the leaf blade length, leaf blade width, and leaf:stem ratio was observed. For the other characteristics, there was a significant linear effect, except for the percentage of dead material. In the dry season, a verified linear response was observed for the chemical characteristics, except neutral detergent insoluble protein. In the rainy season, a linear response was verified for all evaluated characteristics, except acid detergent insoluble protein. Elephant grass BRS Canará for forage production should be cut between 70 and 85 days in the dry season and between 60 and 70 days of regrowth in the rainy season. Objetivou-se determinar a idade de corte ideal do capim-elefante BRS Canará para produção de forragem no período da seca e das águas. O delineamento experimental foi em blocos casualizados, com cinco tratamentos (42, 56, 70, 84 e 98 dias no período da seca e 42, 60, 76, 91 e 105 dias no período das águas) e quatro repetições cada. Foram avaliadas características agronômicas, morfológicas e químicas. No período da seca verificou-se resposta quadrática significativa para altura de planta e comprimento de lâmina foliar. Para as demais características constatou-se resposta linear significativa, com exceção de diâmetro de colmo. No período das águas verificou-se resposta quadrática para comprimento de lâmina foliar, largura de lâmina foliar e relação folha: colmo. Para as demais características verificaram-se efeito linear significativo, com exceção de porcentagem de material senescente. Na seca verificou-se resposta linear das características químicas, exceto para proteína insolúvel em detergente neutro (% MS). No período das águas, verificou-se resposta linear para todas as características avaliadas, exceto para proteína insolúvel em detergente ácido (% PB). O capim-elefante BRS Canará para produção de forragem deve ser cortado entre 70 e 85 dias no período da seca e entre 60 e 70 dias no período das águas.

Muscle foods, particularly fish products are highly exposed to oxidative stress during processing and storage, resulting in oxidative modification of proteins. Protein carbonyls content has been used as one of the measures of oxidative stress. Generally, the resulting carbonylated proteins (CPs) have so far been labeled with 2,4-dinitrophenyl (DNP) hydrazine and detected with anti-DNP antibody. However, the applicability of this method to food samples is limited by its high price, time-consuming procedure and possibility to perform the measurements just on soluble protein fractions. We developed a simpler, faster and cheaper method to assess CP level in muscle foods, including both soluble and insoluble protein fractions, which is based on a direct reaction of protein carbonyls with 7-(diethylamino)coumarin-3-carbohydrazide (CHH). The paper describes a novel technique to label both soluble and insoluble carbonylated proteins with CHH and determine carbonyl content by fluorescence microscopy assay which correlates (R = 0.911) with conventional ELISA method.

Insoluble protein aggregates are the hallmarks of many neurodegenerative diseases. For example, aggregates of TDP-43 occur in nearly all cases of amyotrophic lateral sclerosis (ALS). However, whether aggregates cause cellular toxicity is still not clear, even in simpler cellular systems. We reasoned that deep mutagenesis might be a powerful approach to disentangle the relationship between aggregation and toxicity. We generated >50,000 mutations in the prion-like domain (PRD) of TDP-43 and quantified their toxicity in yeast cells. Surprisingly, mutations that increase hydrophobicity and aggregation strongly decrease toxicity. In contrast, toxic variants promote the formation of dynamic liquid-like condensates. Mutations have their strongest effects in a hotspot that genetic interactions reveal to be structured in vivo, illustrating how mutagenesis can probe the in vivo structures of unstructured proteins. Our results show that aggregation of TDP-43 is not harmful but protects cells, most likely by titrating the protein away from a toxic liquid-like phase., TDP43 aggregates are a hallmark of amyotrophic lateral sclerosis. By using deep mutagenesis to measure the toxicity of more than 50,000 mutations in the prion domain of TDP43, the authors conclude that mutations that increase toxicity promote formation of liquid-like condensates, while aggregation of TDP43 is protective for the cell.

With the advancement of the cutting age, there is an increase in the forage yield of elephant grass (Pennisetum purpureum), but there is also a reduction of the protein levels and digestibility of this forage. This study aimed to identify the ideal cutting age of elephant grass forage (BRS Canará cultivar) to chopped hay production. The experimental design was a randomized block, with five treatments (42, 60, 76, 91 and 105 days of growing) and four replicates. Forage agronomical, morphological and chemical characteristics were evaluated. A linear positive effect of the cutting age was observed on plant height, senescent material and yield of the chopped hay. The leaf percentage and leaf:stem ratio were reduced by the advance in the cutting age. There was also a linear positive effect of the cutting ages on indigestible neutral detergent fiber, neutral detergent fiber corrected for ash and protein and neutral detergent insoluble protein, with increases of 17.13 %, 16.63 % and 20.66 %, respectively. The contents of ashes, crude protein, total digestible nutrients and net lactation energy were reduced with the advance in the forage age. From 76 days, the ashes contents reached values below 9.29 % and the crude protein below 7.16 %. In the same cutting age, the chopped hay yield was 12.91 t ha-1. To improve the hay quality and production, the BRS Canará cultivar must be harvest between 60 and 76 days, when the plants are with 1.20-1.50 m of height, leaf:stem ratio of 0.55-0.44, crude protein of 8.29-7.16 % and indigestible neutral detergent fiber of 26.21-29.06 %. RESUMO Com o avanço da idade de corte, há aumento na produtividade da forragem de capim-elefante (Pennisetum purpureum), mas, também, redução nos teores de proteína e digestibilidade da forrageira. Objetivou-se determinar a época ideal de corte da forragem em capim-elefante (cultivar BRS Canará), para a produção de feno picado. Utilizou-se delineamento experimental em blocos casualizados, com cinco tratamentos (42, 60, 76, 91 e 105 dias de crescimento) e quatro repetições. Foram avaliadas características agronômicas, morfológicas e químicas. Verificou-se efeito linear positivo do momento de colheita sobre a altura da planta, material senescente e produtividade do feno picado. Já a porcentagem de lâminas foliares e a relação folha:colmo reduziram-se, em função da idade de corte. A idade de corte também exerceu efeito linear positivo sobre a fibra em detergente neutro indigestível, fibra em detergente neutro corrigida para cinzas e proteína e proteína insolúvel em detergente neutro, com aumentos de 17,13 %, 16,63 % e 20,66 %, respectivamente. Já os teores de cinzas, proteína bruta, nutrientes digestíveis totais e energia líquida de lactação diminuíram com o avanço da idade da forrageira. A partir de 76 dias, os teores de cinzas atingiram valores abaixo de 9,29 % e a proteína bruta abaixo de 7,16 %. Nesta idade de corte, a produtividade de feno foi de 12,91 t ha-1. Para melhorar a qualidade e a produção de feno, a cultivar BRS Canará deve ser cortada entre 60 e 76 dias, quando as plantas estão com 1,20-1,50 m de altura, relação folha:colmo de 0,55-0,44, proteína bruta de 8,29-7,16 % e fibra em detergente neutro indigestível de 26,21-29,06 %.

While the specificity of protein–lipid interactions is a key feature in the function of biological membranes, studying the specifics of these interactions is challenging because most membrane proteins are insoluble in water due to the hydrophobic nature of their transmembrane domains (TMDs). Here, we introduce a method that overcomes this solubility limitation and identifies the affinity profile of protein TMDs to specific lipid formulations. Using 5 human TMDs as a sample group, our results demonstrate that TMDs are highly selective and that these specific lipid–TMD interactions can involve either a single lipid, or the combination of multiple lipid species.

Summary: Vitamin D has multiple roles, including the regulation of bone and calcium homeostasis. Deficiency of 25-hydroxyvitamin D, the major circulating form of vitamin D, is associated with an increased risk of age-related chronic diseases, including Alzheimer’s disease, Parkinson’s disease, cognitive impairment, and cancer. In this study, we utilized Caenorhabditis elegans to examine the mechanism by which vitamin D influences aging. We found that vitamin-D3-induced lifespan extension requires the stress response pathway genes skn-1, ire-1, and xbp-1. Vitamin D3 (D3) induced expression of SKN-1 target genes but not canonical targets of XBP-1. D3 suppressed an important molecular pathology of aging, that of widespread protein insolubility, and prevented toxicity caused by human β-amyloid. Our observation that D3 improves protein homeostasis and slows aging highlights the importance of maintaining appropriate vitamin D serum levels and may explain why such a wide variety of human age-related diseases are associated with vitamin D deficiency. : Maintenance of protein homeostasis is crucial to cellular health and contributes significantly to the lifespan of organisms. Mark et al. demonstrate that vitamin D supplementation promotes protein homeostasis and slows aging in the nematode, C. elegans. These findings identify a mechanism by which vitamin D influences aging. Keywords: Ceanorhabditis elegans, vitamin D, lifespan. aging, insoluble protein, SKN-1, XBP-1, IRE-1, proteostasis, protein aggregation, Alzheimer’s disease

In the 1940s, research by Baker established fundamental aspects of the breadmaking process while Finney showed that differences of quality resided in the gluten proteins of flour. Different lipid components have been found to affect loaf volume and texture but variation in lipid composition of current wheat varieties does not account for significant quality differences. Fundamental studies by Tipples and Kilborn showed that optimum dough development had two requirements – a critical mixing intensity and a critical amount of imparted energy. Expansion of a fermenting dough depends on two stabilizing mechanisms resulting from the gluten/starch matrix and the liquid lamellae surrounding the gas bubbles. In the 1970s, wheat cultivars were developed with poor baking properties, related to a dearth of insoluble protein, shown to comprise glutenins of large molecular size. Research by Payne and co-workers revealed that dough strength could be related to glutenin subunits and the genes controlling their synthesis. This has opened up new directions for determining composition-quality relationships and, in turn, has enabled strategies to be devised for improving quality based on wheat genetic composition.

This research is focused on proteolysis and total antioxidant capacity of proteins of white brined cheese prepared from overheated (90 degrees C, 10 minutes) cow milk. White brined cow cheese of overheated milk went through specific proteolytic changes during ripening that were result of high level of whey proteins incorporated into the gel matrix. Specificity was reflected through the relatively low level of soluble nitrogen fractions, the intensive and continual decrease of alpha(s)-caseins up to 15.42 % of initial content, slow degradation of beta-casein throughout the whole ripening period and high level of proteolytic products tightly bounded into gel matrix. Strong negative correlations (-0.97, -0.98 and -0.91; p lt 0.05) between ripening time and resudual alpha(s)-caseins, beta-casein and low molecular weight products were observed. Proteolysis also affected the total antioxidant capacity of both water soluble and water insoluble nitrogen fractions, but to different extents and with different trends. Total antioxidant capacity of water insoluble fraction increased slowly during the whole ripening period, wherease significant improvement of total antioxidant capacity of water soluble fraction started after 30 days of ripening. These findings could be useful for better understanding and control of the white brined cow cheese production.

Alzheimer disease (AD) is the most devastating and widespread neurodegenerative disorder around the globe. Insoluble protein aggregates originate from beta amyloid peptides is the hall mark of the disease. These peptides are produced via cleavage of Amyloid Precursor Protein (APP) by Bace1 enzyme. Therefore, finding a drug that will specifically inhibit this enzyme is one of the attractive approaches for AD treatment. Here, we assessed the affinity of 323 inhibitors that have co-crystallized with Bace1 enzyme towards two structurally related enzymes Bace2 and CatD by two different docking software Glide-QPLD and GOLD. Based on two docking results, 3% of all ligands have been found to prefer Bace1 enzyme over the other two enzymes. Interestingly, all of these ligands have –NH2 functional groups interacting with the catalytic dyad (Asp80 and Asp 276), which suggests the important role of the –NH 2 adduct in designing Bace1 specific inhibitors.