Analysis of Bacterial DNA-Binding Protein from Starved Cells

Our experts have a keen interest in the study of liquid-liquid and liquid-solid phase transitions in bacterial cells. We have cutting-edge super-resolution imaging or infinite diffraction microscopy apparently combined with single molecule trafficking methods, and computer analysis platforms to analyze key microbial biomolecular condensates undergoing LLPS, as well as the formation and organization of biomolecular condensates within the intracellular space. Here, CD BioSciences is committed to analyzing intrinsically disordered bacterial DNA-binding protein from starved cells (Dps).

Introduction of Bacterial DNA-Binding Protein from Starved Cells

The ability to adapt to environmental changes is one of the key determinants of an organism's adaptability. Bacteria have evolved multiple ways to survive and thrive under stressful conditions, ranging from extreme measures (such as spore production) to specialized stress-mediated protein expression. DNA-binding protein from starved cells (Dps) is a critical protein in stress survival and is highly conserved in more than 47 bacterial species. The presence of Dps enhances bacterial survival under many different stresses, including starvation, heat shock, oxidative stress, and iron overexposure. These protective effects of Dps expression may be due to its dual biochemical functions, DNA binding and iron oxidase activity. However, the molecular mechanisms and physiological consequences of these activities have not been fully elucidated. Studies suggest that the E. coli nucleoid protein Dps may be organized by liquid-liquid phase separation (LLPS).

Fig. 1. Structural organization of Dps proteins. (Chiancone E, et al., 2010)

Customized Services

One of the adaptive strategies for changing environmental conditions in bacterial cells involves the condensation of DNA/iron oxidase complexes with Dps. The structures of Dps all contain intrinsically disordered regions. Our technical team has successfully established E. coli as a model system to identify the structure and key residues of Dps and to create Dps variants and mutant E. coli strains.

CD BioSciences provides cutting-edge in vitro and computer-based platforms to analyze the structural dynamics and biochemical functions of intrinsically disordered Dps in bacteria. Our technology platforms are as follows:

• We provide structured illumination microscopy for imaging overexpression of fluorescent Dps protein constructs, allowing direct observation of the morphology of Dps condensates within live E. coli cells. We aim to verify the presence of fluorescent Dps proteins by using their visualization.
• We offer different imaging modalities to characterize Dps LLPS in vitro, including bright-field imaging, differential interference contrast imaging, phase contrast imaging, wide-field fluorescence microscopy, and confocal microscopy imaging. In addition, we have fluorescence recovery after photobleaching (FRAP) technique to analyze the material properties of Dps, including condensate-condensate fusion, condensate dropout, condensate surface wetting, and fluorescence recovery, etc.
• We offer turbidity as a metric for condensate formation in LLPS, allowing full analysis of intermolecular interactions in Dps phase separation.
• Our experts are committed to developing therapeutic Dps targets to solve many antibiotic-related bioengineering and medical problems.

CD BioSciences offers a simple modular biomolecular platform to characterize the LLPS of intrinsically disordered Dps. We aim to help our clients understand the unique role of Dps proteins in bacterial resistance to general and specific stress conditions from a molecular perspective, and develop therapeutic targets. If you have any special requirements for our services, please feel free to contact us.