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Biology subjects

Garay, E.

Publications and source records attributed to Garay, E..

4 recordsLinked to original sources

ProtoPure: Literature Mining and Analysis of Protein Purification Protocols at Proteome Scale

Protein purification is a fundamental step for analyzing individual proteins and protein complexes and identifying interactions with other proteins, DNA, or RNA. A variety of protein purification strategies exist to address desired scale, throughput, and downstream applications. However, selecting an effective experimental procedure remains a largely empirical process that a quick literature review can help guide. Researchers typically survey on the order of a dozen publications for proteins with similar biochemical properties, assessing conditions on a case-by-case basis, leaving no systematic framework for comparison and decision-making at the scale of entire protein families. As protein engineering and design evolve, identifying effective purification conditions remains critical for obtaining sufficient quantities of properly folded and functional proteins. To address this gap, we developed ProtoPure, an LLM-based workflow and database that automates the extraction, standardization and analysis of protein purification conditions from peer-reviewed literature at scale. Given a user-defined UniProt table-enabling flexible selection of proteins by family, organism, or proteome. ProtoPure retrieves associated publications, classifies them based on the presence of purification protocols, and extracts a standardized set of 12 purification parameters (e.g., buffer composition, elution conditions). By transforming dispersed experimental descriptions into a structured resource, ProtoPure makes it possible to compare purification strategies across thousands of proteins. The resulting dataset revealed quantitative convergence toward a limited set of recombinant expression practices, with BL21(DE3)-based systems dominating protein production and recurrent combinations of expression host, inducer, growth medium, and plasmid occurring substantially more frequently than expected by chance. In contrast, downstream purification parameters exhibited greater diversity, revealing a workflow architecture that combines standardized expression infrastructure with more flexible biochemical components.

biophysics↗

Bats from the Colombian Caribbean Reveal a new subtype of Influenza A (H18N12)

Influenza viruses have an excellent capacity for mutation and adaptation in mammalian hosts, which makes them viruses of medical and veterinary importance. Influenzaviruses have been studied mainly in birds but minor in bats. It is unknown whether Chiroptera are reservoirs of influenza viruses. However, circulation in bats showed molecular divergence from H17N10 (Guatemala) and H18N11 (Peru), and they were designated as new subtypes. The study aimed to characterize the influenza A virus detected in the fishing bat Noctilio albiventris. A surveillance study of pathogens of public health interest was carried out; rectal samples were taken from four fishing bats (N. albiventris) captured in Talaigua Nuevo, Bolivar, Colombia. The samples were sequenced by NGS using DNBseq (MGI-G50(R)) and analyzed with bioinformatics tools. Eight viral contigs associated with the Orthomyxoviridae family were obtained. The identified segments showed around 90% similarity with H18N11, except for the neuraminidase (N). The phylogenetic analysis of the N protein showed the appearance of a basal branch to the N11 subtype, and the molecular clock indicates that it does not share a recent common ancestor. 3D modeling indicates that the N protein of N. albiventris presents three mutations (K363R, T242K, and I139V) near the hypothetical active site of the protein. These mutations potentially increase the interaction with the HLA-DR of bats, which could have significant implications for the viruss behavior. The phylogenetic, evolutionary, and antigenic divergence of the N protein of N. albiventris suggests a new subtype called H18N12. Its role as a pathogen must be studied.

bioinformatics↗

A Competitive-Aging Method for Quantitative Genetic Analysis of the Chronological Lifespan of Saccharomyces cerevisiae

The chronological lifespan of budding yeast is a model of aging and age-related diseases. This paradigm has recently allowed genome-wide screening of genetic factors underlying post-mitotic viability in a simple unicellular system, which underscores its potential to provide a comprehensive view of the aging process. However, results from different large-scale studies show little overlap and typically lack quantitative resolution to derive interactions among different aging factors. We previously introduced a sensitive, parallelizable approach to measure the chronological-lifespan effects of gene deletions based on the competitive aging of fluorescence-labeled strains. Here, we present a thorough description of the method, including an improved multiple-regression model to estimate the association between death rates and fluorescent signals, which accounts for possible differences in growth rate and experimental batch effects. We illustrate the experimental procedure--from data acquisition to calculation of relative survivorship--for ten deletion strains with known lifespan phenotypes, which is achieved with high technical replicability. We apply our method to screen for gene-drug interactions in an array of yeast deletion strains, which reveals a functional link between protein glycosylation and lifespan extension by metformin. Competitive-aging screening coupled to multiple-regression modeling provides a powerful, straight-forward way to identify aging factors in yeast and their interactions with pharmacological interventions.

genetics↗

Genome-Wide Mechanisms of Lifespan Extension by Dietary Restriction in Yeast

Dietary restriction is arguably the most promising non-pharmacological intervention to extend human life and health span. Yet, only few genetic regulators mediating the cellular response to dietary restriction are known, and the question remains which other transcription factors and regulatory pathways are involved. To gain a comprehensive view of how lifespan extension under dietary restriction is elicited, we screened the chronological lifespan of most gene deletions of Saccharomyces cerevisiae under two dietary regimens, restricted and non-restricted. We identified 472 mutants with enhanced or diminished extension of lifespan by dietary restriction. Functional analysis of such dietary-restriction genes revealed novel processes underlying longevity specifically by dietary restriction. Importantly, this set of genes allowed us to generate a prioritized catalogue of transcription factors orchestrating the dietary-restriction response, which underscored the relevance of cell-cycle arrest control as a key mechanism of chronological longevity in yeast. We show that the transcription factor Ste12 is needed for full lifespan extension and cell-cycle arrest in response to nutrient limitation; linking the pheromone/invasive growth pathway with cell survivorship. Strikingly, STE12 overexpression was sufficient to extend chronological lifespan under non-restricted conditions. Our global picture of the genetic players of longevity by dietary restriction highlights intricate regulatory cross-talks in aging cells.

genetics↗