25 research outputs found

    Proteome-wide dataset supporting the study of ancient metazoan macromolecular complexes.

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    Our analysis examines the conservation of multiprotein complexes among metazoa through use of high resolution biochemical fractionation and precision mass spectrometry applied to soluble cell extracts from 5 representative model organisms Caenorhabditis elegans, Drosophila melanogaster, Mus musculus, Strongylocentrotus purpuratus, and Homo sapiens. The interaction network obtained from the data was validated globally in 4 distant species (Xenopus laevis, Nematostella vectensis, Dictyostelium discoideum, Saccharomyces cerevisiae) and locally by targeted affinity-purification experiments. Here we provide details of our massive set of supporting biochemical fractionation data available via ProteomeXchange (PXD002319-PXD002328), PPIs via BioGRID (185267); and interaction network projections via (http://metazoa.med.utoronto.ca) made fully accessible to allow further exploration. The datasets here are related to the research article on metazoan macromolecular complexes in Nature [1]

    Panorama of ancient metazoan macromolecular complexes.

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    Macromolecular complexes are essential to conserved biological processes, but their prevalence across animals is unclear. By combining extensive biochemical fractionation with quantitative mass spectrometry, here we directly examined the composition of soluble multiprotein complexes among diverse metazoan models. Using an integrative approach, we generated a draft conservation map consisting of more than one million putative high-confidence co-complex interactions for species with fully sequenced genomes that encompasses functional modules present broadly across all extant animals. Clustering reveals a spectrum of conservation, ranging from ancient eukaryotic assemblies that have probably served cellular housekeeping roles for at least one billion years, ancestral complexes that have accrued contemporary components, and rarer metazoan innovations linked to multicellularity. We validated these projections by independent co-fractionation experiments in evolutionarily distant species, affinity purification and functional analyses. The comprehensiveness, centrality and modularity of these reconstructed interactomes reflect their fundamental mechanistic importance and adaptive value to animal cell systems

    Atividade de isolados bacterianos solubilizadores de fosfato na presença de formulações comerciais de Glyphosate Activity of phosphate solubilizing bacterial isolates in the presence of commercial glyphosate formulations

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    Objetivou-se com este trabalho avaliar os efeitos da aplicação de formulações comerciais de glyphosate (Roundup Transorb®, Zapp QI®, Roundup NA® e Scout®) na capacidade de dois isolados bacterianos (To 11 e To 66) em solubilizar diferentes fontes inorgânicas de fosfato. A atividade de solubilização de fosfato inorgânico dos isolados bacterianos foi avaliada em três fontes de fosfato inorgânico (fosfato de cálcio, de alumínio e de ferro), na presença de diferentes formulações de glyphosate na concentração de 60 mg L-1 do equivalente ácido e tratamento controle sem adição dos herbicidas. Os efeitos das formulações de glyphosate foram diferentes para cada isolado. As formulações Roundup Transorb® e Zapp QI® provocaram redução na capacidade de solubilização do isolado To 66, enquanto a formulação Scout® aumentou essa capacidade. Por sua vez, o isolado To 11 não teve sua capacidade de solubilização afetada na presença das formulações avaliadas. Em média, as formulações Roundup NA® e Scout® não alteraram a capacidade de solubilização dos isolados, ao passo que os herbicidas Roundup Transorb® e Zapp QI® reduziram essa capacidade de solubilização.<br>The objective of this work was to evaluate the effect of applying different commercial formulations of glyphosate (Roundup Transorb®, Zapp QI®, Roundup NA® e Scout®) on the ability of two bacterial isolates (To 11 and To 66) to solubilize different inorganic phosphates. The ability to solubilize bacterial isolate phosphate was evaluated in relation to three inorganic phosphate sources (calcium, aluminum, and iron phosphates) in the presence of different commercial formulations of glyphosate, at the concentration of 60 mg L-1 acid equivalent, and control treatment without the addition of the herbicides. The effects of the commercial formulations of glyphosate on each bacterial isolate were distinct. Roundup Transorb and Zapp QI led to a significant decrease in the phosphate-solubilizing activity of To 66, while the reverse was observed for Scout. To 11 did not show significant differences in phosphate solubilization in the presence of the commercial formulations tested. Roundup NA and Scout did not change the ability to solubilize phosphate of bacterial isolates, while Roundup Transorb and Zapp QI led to a decrease in the solubilizing activity

    A Map of Human Mitochondrial Protein Interactions Linked to Neurodegeneration Reveals New Mechanisms of Redox Homeostasis and NF-\u3baB Signaling

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    Mitochondrial protein (MP) dysfunction has been linked to neurodegenerative disorders (NDs); however, the discovery of the molecular mechanisms underlying NDs has been impeded by the limited characterization of interactions governing MP function. Here, using mass spectrometry (MS)-based analysis of 210 affinity-purified mitochondrial (mt) fractions isolated from 27 epitope-tagged human ND-linked MPs in HEK293 cells, we report a high-confidence MP network including 1,964 interactions among 772 proteins (> 90% previously unreported). Nearly three-fourths of these interactions were confirmed in mouse brain and multiple human differentiated neuronal cell lines by primary antibody immunoprecipitation and MS, with many linked to NDs and autism. We show that the SOD1-PRDX5 interaction, critical for mt redox homeostasis, can be perturbed by amyotrophic lateral sclerosis-linked SOD1 allelic variants and establish a functional role for ND-linked factors coupled with I kappa B epsilon in NF-kappa B activation. Our results identify mechanisms for ND-linked MPs and expand the human mt interaction landscape
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