It has remained unknown how cells reduce cystine taken up from the extracellular space, which is a required step for further utilization of cysteine in key processes such as protein or glutathione synthesis. Here, we show that the thioredoxin-related protein of 14 kDa (TRP14, encoded by TXNDC17 ) is the rate-limiting enzyme for intracellular cystine reduction. When TRP14 is genetically knocked out, cysteine synthesis through the transsulfuration pathway becomes the major source of cysteine in human cells, and knockout of both pathways becomes lethal in C. elegans subjected to proteotoxic stress. TRP14 can also reduce cysteinyl moieties on proteins, rescuing their activities as here shown with cysteinylated peroxiredoxin 2. Txndc17 knockout mice were, surprisingly, protected in an acute pancreatitis model, concomitant with activation of Nrf2-driven antioxidant pathways and upregulation of transsulfuration. We conclude that TRP14 is the evolutionarily conserved enzyme principally responsible for intracellular cystine reduction in C. elegans , mice, and humans.

Leishmania infantum causes visceral leishmaniasis (kala-azar), the most severe form of leishmaniasis, which is lethal if untreated. A few years ago, the re-sequencing and de novo assembling of the L. infantum (JPCM5 strain) genome was accomplished, and now we aimed to describe and characterize the experimental proteome of this species. In this work, we performed a proteomic analysis from axenic cultured promastigotes and carried out a detailed comparison with other Leishmania experimental proteomes published to date. We identified 2352 proteins based on a search of mass spectrometry data against a database built from the six-frame translated genome sequence of L. infantum. We detected many proteins belonging to organelles such as glycosomes, mitochondria, or flagellum, as well as many metabolic enzymes and many putative RNA binding proteins and molecular chaperones. Moreover, we listed some proteins presenting post-translational modifications, such as phosphorylations, acetylations, and methylations. On the other hand, the identification of peptides mapping to genomic regions previously annotated as non-coding allowed for the correction of annotations, leading to the N-terminal extension of protein sequences and the uncovering of eight novel protein-coding genes. The alliance of proteomics, genomics, and transcriptomics has resulted in a powerful combination for improving the annotation of the L. infantum reference genome.

The impact of the world’s growing population on food systems and the role of dietary patterns in the management of non-communicable diseases underscore the need to explore sustainable and dietary protein sources. Although microalgae have stood out as alternative sources of proteins and bioactive peptides, some species such as Nannochloropsis gaditana remain unexplored. This study aimed to characterize N. gaditana’s proteome and evaluate its potential as a source of bioactive peptides by using an in silico approach. A total of 1955 proteins were identified and classified into functional groups of cellular components, molecular functions, and biological processes. In silico gastrointestinal digestion of identified proteins demonstrated that 202 hydrophobic and low-molecular-size peptides with potential bioactivity were released. Among them, 27 exhibited theorical antioxidant, antihypertensive, antidiabetic, anti-inflammatory, and/or antimicrobial activities. Seven of twenty-seven peptides showed ≥20% intestinal absorption, suggesting potential systemic effects, while the rest could act at local level. Molecular docking demonstrated strong affinities with key enzymes such as MPO, ACE, and DPPIV. Resistance to the digestion, capacity to be absorbed, and multifunctionality were demonstrated for peptide FIPGL. This study highlights N. gaditana’s potential as a sustainable source of novel potential bioactive peptides with promising local and systemic biological effects.

The innate and adaptative immune systems are involved in the regulation of inflammatory and oxidative processes and mediators such as reactive oxygen species (ROS) and nitric oxide (NO). The exacerbated action of these players results in an oxidative stress status and chronic inflammation, which is responsible for the development of non-communicable diseases (NCDs). By modulating these mediators, bioactive compounds in food can exert a key role in the prevention of several NCDs. Among these compounds, soybean proteins and peptides such as lunasin have been considered to be among the most promising. The aim of this study was to obtain and characterize a soluble protein-enriched extract from a commercial soybean protein isolate and fractionate it into different fractions through ultrafiltration. Their antioxidant and immunomodulatory properties were then evaluated using biochemical and cell models. A total of 535 proteins (from 282 protein groups) were identified in the extract, in which the presence of the peptide lunasin was confirmed. The enrichment of this peptide was achieved in the 3–10 kDa fraction. The protective effects against the oxidative stress induced by LPS in the macrophage model could have been mediated by the radical scavenging capacity of the peptides present in the soybean samples. Under basal conditions, the extract and its ultrafiltered fractions activated macrophages and induced the release of NO. However, under challenged conditions, the whole extract potentiated the NO-stimulating effects of LPS, whereas the fraction containing 3–10 kDa peptides, including lunasin, counteracted the LPS-induced NO increase. Our findings suggest a promising role of soybean protein as an ingredient for functional foods and nutraceuticals aimed at promoting health and preventing oxidative stress and/or immune-alteration-associated diseases.

The high-throughput proteomics data generated by increasingly more sensible mass spectrometers greatly contribute to our better understanding of molecular and cellular mechanisms operating in live beings. Nevertheless, proteomics analyses are based on accurate genomic and protein annotations, and some information may be lost if these resources are incomplete. Here, we show that most proteomics data may be recovered by interconnecting genomics and proteomics approaches (i.e., following a proteogenomic strategy), resulting, in turn, in an improvement of gene/protein models. In this study, we generated proteomics data from Leishmania donovani (HU3 strain) promastigotes that allowed us to detect 1908 proteins in this developmental stage on the basis of the currently annotated proteins available in public databases. However, when the proteomics data were searched against all possible open reading frames existing in the L. donovani genome, twenty new protein-coding genes could be annotated. Additionally, 43 previously annotated proteins were extended at their N-terminal ends to accommodate peptides detected in the proteomics data. Also, different post-translational modifications (phosphorylation, acetylation, methylation, among others) were found to occur in a large number of Leishmania proteins. Finally, a detailed comparative analysis of the L. donovani and Leishmania major experimental proteomes served to illustrate how inaccurate conclusions can be raised if proteomes are compared solely on the basis of the listed proteins identified in each proteome. Finally, we have created data entries (based on freely available repositories) to provide and maintain updated gene/protein models. Raw data are available via ProteomeXchange with the identifier PXD051920.

Chagas disease is caused by the intracellular protozoan parasite Trypanosoma cruzi. This disease affects mainly rural areas in Central and South America, where the insect vector is endemic. However, this disease has become a world health problem since migration has spread it to other continents. It is a complex disease with many reservoirs and vectors and high genetic variability. One of the host proteins involved in the pathogenesis is SLAMF1. This immune receptor acts during the infection of macrophages controlling parasite replication and thus affecting survival in mice but in a parasite strain-dependent manner. Therefore, we studied the role of SLAMF1 by quantitative proteomics in a macrophage in vitro infection and the different responses between Y and VFRA strains of Trypanosoma cruzi. We detected different significant up- or downregulated proteins involved in immune regulation processes, which are SLAMF1 and/or strain-dependent. Furthermore, independently of SLAMF1, this parasite induces different responses in macrophages to counteract the infection and kill the parasite, such as type I and II IFN responses, NLRP3 inflammasome activation, IL-18 production, TLR7 and TLR9 activation specifically with the Y strain, and IL-11 signaling specifically with the VFRA strain. These results have opened new research fields to elucidate the concrete role of SLAMF1 and discover new potential therapeutic approaches for Chagas disease.