Analysis of Biomolecular Condensates in Microbial Cells
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Analysis of Biomolecular Condensates in Microbial Cells

In eukaryotic cells, many protein systems can spontaneously organize into macromolecular condensates formed by liquid-liquid phase separation (LLPS). These organelles without membranes provide a strategy for organizing cells without physical barriers while allowing for the dynamic, reactive organization of cells. With advances in fluorescence imaging and single-molecule microscopy techniques, the discovery of biomolecular condensates in bacteria and protists is rapidly increasing. The structure, function, and potential applications of these microbial condensates are currently receiving extensive attention. Scientists have been able to study the characteristics of LLPS in vitro and in vivo by utilizing the latest imaging, structural and computational methods. However, in-depth studies of LLPS in relatively small prokaryotic cells remain technically challenging, limiting our understanding of the molecular basis and biological functions of microbial compartmentalization.

Fig. 1. Proposed phase transitions in bacterial cells.Fig. 1. Proposed phase transitions in bacterial cells. (Sołtys K, et al., 2022)

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Studies have shown that bacterial cytoplasm displays glass-forming fluid properties and can solidify into soft glass-like forms, depending on metabolism, composition size, and non-site blocking interactions. Our lab has cutting-edge modern microscopy techniques to analyze biomolecular condensates in bacteria and protists, including the formation of phase-separated condensates in E. coli, Bacillus subtilis, Bacillus crescentus, etc. Our experts are committed to analyzing their biogenesis and biological function. In addition, we are exploring the molecular syntax of these biomolecular condensates to rationalize the design of synthetic condensates in mammalian cells.

CD BioSciences offers comprehensive biomolecular condensate analysis services in microbial cells to test the breadth of bacterial utilization of LLPS, examine its impact on enzyme kinetics and design synthetic membrane-free organelles. Our lab can characterize more than 10 bacterial LLPS systems, as following:

LLPS Systems in Prokaryotic Microbes Representative Species
Polar organizing protein Z (PopZ) Caulobacter crescentus
Bacterial ribonucleoprotein bodies (BR-bodies) Caulobacter crescentus
Filamentous temperature-sensitive Z (FtsZ) Escherichia coli
Carboxysomes Synechococcus elongatus
ParABS system Escherichia coli
RNA polymerase (RNAP) Escherichia coli
Single-stranded DNA-binding protein (SSB) Escherichia coli
ATP-binding cassette transporters Mycobacterium tuberculosis
Polyphosphate granules Pseudomonas aeruginosa
DNA-binding protein from starved cells (Dps) Escherichia coli

Our Technology Platform for Studying Phase-Separated Condensates in Prokaryotic Systems

We offer super-resolution imaging or infinite diffraction display combined with single molecule trafficking methods for the study of cells in LLPS and liquid condensates in prokaryotic systems, allowing for quantitative analysis.

  • The single molecule localization microscopy (SMLM) technology platform includes Stochastic Optical Reconstruction Microscopy (STORM), Photo Activated Localization Microscopy (PALM) and its various modifications. This technology platform allows precise determination of molecular trajectories and provides information about the dynamics of molecules in a condensate.
  • Infinite diffraction microscopy technology platform. This technique has high throughput and is not susceptible to reconstruction artifacts.
  • Computer-based analysis platform. We offer a number of LLPS prediction tools based on different algorithms to analyze bacterial proteins with LLPS and condensate formation tendencies.

Advantages of Studying Bacterial and Protists Condensates

  • They are ideal for in vitro recombination because of their relatively simple composition and help establish a direct link between phase separation and biological function.
  • Bacteria and protists span the tree of life and exhibit a wide range of metabolic and ecological strategies. This diversity allows for comparative analysis of condensate structure and function.
  • Bacterial and protozoan condensates provide a powerful substrate for synthetic biology.

CD BioSciences aims to help customers characterize biomolecular condensates in microorganisms, and analyze their formation and organization. Our services will open up more possibilities for future drug discovery. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together on your attractive projects.

Reference

  1. Sołtys K, Tarczewska A, et al. (2022) Getting Closer to Decrypting the Phase Transitions of Bacterial Biomolecules. Biomolecules. 12(7):907.
For research use only, not intended for any clinical use.
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