Decoding the stoichiometric composition and organisation of bacterial metabolosomes

• Published in: Nature communications Vol. 11; no. 1; p. 1976
• Main Authors: Yang, Mengru, Simpson, Deborah M., Wenner, Nicolas, Brownridge, Philip, Harman, Victoria M., Hinton, Jay C. D., Beynon, Robert J., Liu, Lu-Ning
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24.04.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 14/19; 14/28; 631/1647/296; 631/326/41/2531; 631/45/475; 631/80/642; 82/58; Bacteria; Bacterial Proteins - metabolism; Biodegradation; Biotechnology; Catalysis; Composition; Cytoplasm - metabolism; Cytosol; Gene Expression Regulation, Bacterial; Green Fluorescent Proteins - metabolism; Humanities and Social Sciences; Mass Spectrometry; Mass spectroscopy; Metabolomics; multidisciplinary; Mutation; Organizational aspects; Pathogenesis; Propylene Glycol - pharmacology; Protein Conformation; Proteins; Proteomics; Salmonella; Salmonella enterica - genetics; Salmonella enterica - metabolism; Salmonella typhimurium - genetics; Salmonella typhimurium - metabolism; Science; Science (multidisciplinary); Scientific imaging; Self-assembly; Spectroscopy; Stoichiometry; Structure-function relationships; Synthetic Biology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-15888-4

Some enteric bacteria including Salmonella have evolved the propanediol-utilising microcompartment (Pdu MCP), a specialised proteinaceous organelle that is essential for 1,2-propanediol degradation and enteric pathogenesis. Pdu MCPs are a family of bacterial microcompartments that are self-assembled from hundreds of proteins within the bacterial cytosol. Here, we seek a comprehensive understanding of the stoichiometric composition and organisation of Pdu MCPs. We obtain accurate stoichiometry of shell proteins and internal enzymes of the natural Pdu MCP by QconCAT-driven quantitative mass spectrometry. Genetic deletion of the major shell protein and absolute quantification reveal the stoichiometric and structural remodelling of metabolically functional Pdu MCPs. Decoding the precise protein stoichiometry allows us to develop an organisational model of the Pdu metabolosome. The structural insights into the Pdu MCP are critical for both delineating the general principles underlying bacterial organelle formation, structural robustness and function, and repurposing natural microcompartments using synthetic biology for biotechnological applications. Enteric pathogens such as Salmonella depend on propanediol-utilising microcompartments (Pdu MCP), which self-assemble from cytosolic proteins. Using mass spectrometry-based absolute quantification, the authors here define the protein stoichiometry and propose an organizational model of a Salmonella Pdu MCP.