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Ultradynamic protein assemblies and network channeling

ABSTRACT: Whereas a classical view sustains that sequential metabolic reactions and other complex step-wise processes can be efficiently executed by proteins dispersed in a crowded intracellular environment, recent evidence in some metabolic pathways points to the notion that sequential enzymes would be somehow organized in protein assemblies. However, the existence of stable complexes holding a large number of different enzymes by multiple keylock interactions is rather exceptional. Based on fluorescence fluctuation analysis, we have found that protein pairs performing enzymatic steps of glycolysis display strong spatiotemporal coincidence compared to unrelated proteins acting in other pathways. Notably, spatiotemporal coincidence inversely correlated with biochemical distance within the glycolytic pathway. These protein assemblies, which we propose to name protein flocks, display association and dissociation dynamics under the millisecond, well beyond those displayed by membrane-less organelles. To confirm and characterize the compositional specificity of protein flocks, we have applied proximity-labeling and MS assays to glycolysis proteins.

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ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Martí Aldea

LAB HEAD: Marti Aldea

PROVIDER: PXD065061 | Pride | 2026-05-04

REPOSITORIES: Pride

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Project description:Microbial communities often undergo intricate compositional changes yet also maintain stable coexistence of diverse species. The mechanisms underlying long-term coexistence remain unclear, as system-wide studies have been largely limited to engineered communities, ex situ adapted cultures, or synthetic assemblies. Here we show how kefir, a natural milk-fermenting community of prokaryotes and yeasts, realises stable coexistence through spatiotemporal orchestration of species and metabolite dynamics. During milk fermentation, kefir grains (a polysaccharide matrix synthesized by kefir microbes) grow in mass but remain unchanged in composition. In contrast, the milk is colonized in a sequential manner in which early members open metabolic niches for followers. Through metabolomics and large-scale mapping of inter-species interactions, we show how microbes poorly suited for milk survive in, and even dominate the community, through metabolic cooperation and uneven partitioning between grain and milk. Overall, our findings reveal how inter-species interactions partitioned in space and time lead to stable coexistence. Linked metabolomics studies:<a href='https://www.ebi.ac.uk/metabolights/MTBLS1829' rel='noopener noreferrer' target='_blank'>MTBLS1829</a> Kefir fermentation curve (FIA-MS)<a href='https://www.ebi.ac.uk/metabolights/MTBLS1830' rel='noopener noreferrer' target='_blank'>MTBLS1830</a> Interaction between the kefir isolates Lactococcus lactis and Acetobacter fabarum (GC-MS) Linked cross omic data:Genomes of isolated kefir species are available in the <a href='https://www.ncbi.nlm.nih.gov/' rel='noopener noreferrer' target='_blank'>NCBI database</a> under the accession: PRJNA375758 (bioproject ID: 375758).Metatranscriptomic sequencing reads can be accessed from <a href='www.ebi.ac.uk/ena' rel='noopener noreferrer' target='_blank'>ENA</a> under the project id PRJEB37001.Genome-scale metabolic models for kefir bacteria can be found at github.com/cdanielmachado/kefir_models.

Project description:Microbial communities often undergo intricate compositional changes yet also maintain stable coexistence of diverse species. The mechanisms underlying long-term coexistence remain unclear, as system-wide studies have been largely limited to engineered communities, ex situ adapted cultures, or synthetic assemblies. Here we show how kefir, a natural milk-fermenting community of prokaryotes and yeasts, realizes stable coexistence through spatiotemporal orchestration of species and metabolite dynamics. During milk fermentation, kefir grains (a polysaccharide matrix synthesized by kefir microbes) grow in mass but remain unchanged in composition. In contrast, the milk is colonized in a sequential manner in which early members open metabolic niches for followers. Through metabolomics and large-scale mapping of inter-species interactions, we show how microbes poorly suited for milk survive in — and even dominate — the community, through metabolic cooperation and uneven partitioning between grain and milk. Overall, our findings reveal how inter-species interactions partitioned in space and time lead to stable coexistence. Linked metabolomics studies:<a href='https://www.ebi.ac.uk/metabolights/MTBLS1823' rel='noopener noreferrer' target='_blank'>MTBLS1823</a> Spent-medium assay, measured untargeted (FIA-qTOF) and targeted (HILIC, LC-MS)<a href='https://www.ebi.ac.uk/metabolights/MTBLS1829' rel='noopener noreferrer' target='_blank'>MTBLS1829</a> Kefir fermentation curve (FIA-MS) Linked cross omic data:Genomes of isolated kefir species are available in the <a href='https://www.ncbi.nlm.nih.gov/' rel='noopener noreferrer' target='_blank'>NCBI database</a> under the accession: PRJNA375758 (bioproject ID: 375758).Metatranscriptomic sequencing reads can be accessed from <a href='https://www.ebi.ac.uk/metabolights/editor/www.ebi.ac.uk/ena' rel='noopener noreferrer' target='_blank'>ENA</a> under the project id PRJEB37001.Genome-scale metabolic models for kefir bacteria can be found at github.com/cdanielmachado/kefir_models.

Project description:Microbial communities often undergo intricate compositional changes yet also maintain stable coexistence of diverse species. The mechanisms underlying long-term coexistence remain unclear, as system-wide studies have been largely limited to engineered communities, ex situ adapted cultures, or synthetic assemblies. Here we show how kefir, a natural milk-fermenting community of prokaryotes and yeasts, realises stable coexistence through spatiotemporal orchestration of species and metabolite dynamics. During milk fermentation, kefir grains (a polysaccharide matrix synthesized by kefir microbes) grow in mass but remain unchanged in composition. In contrast, the milk is colonized in a sequential manner in which early members open metabolic niches for followers. Through metabolomics and large-scale mapping of inter-species interactions, we show how microbes poorly suited for milk survive in, and even dominate the community, through metabolic cooperation and uneven partitioning between grain and milk. Overall, our findings reveal how inter-species interactions partitioned in space and time lead to stable coexistence. Linked metabolomics studies:<a href='https://www.ebi.ac.uk/metabolights/MTBLS1823' rel='noopener noreferrer' target='_blank'>MTBLS1823</a> Spent-medium assay, measured untargeted (FIA-qTOF) and targeted (HILIC, LC-MS)<a href='https://www.ebi.ac.uk/metabolights/MTBLS1830' rel='noopener noreferrer' target='_blank'>MTBLS1830</a> Interaction between the kefir isolates Lactococcus lactis and Acetobacter fabarum (GC-MS) Linked cross omic data sets:Genomes of isolated kefir species are available in the NCBI database under the accession: PRJNA375758 (bioproject ID: 375758).Metatranscriptomic sequencing data associated with this study are available in the European Nucleotide Archive (ENA): accession number <a href='https://www.ebi.ac.uk/ena/browser/view/PRJEB37001' rel='noopener noreferrer' target='_blank'>PRJEB37001</a>.Genome-scale metabolic models for kefir bacteria can be found at github.com/cdanielmachado/kefir_models.

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Ultradynamic protein assemblies and network channeling

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