Microbial species have developed multiple mechanisms to mitigate damage caused by acidic environments, including maintenance of pH homeostasis, cell membrane integrity and fluidity, metabolic regulation, and macromolecular repair. Maintenance of pH homeostasis is critical for microbial survival in acidic environments. Here, CD BioSciences offers a specialized microbial vesicle acidification assessment service to analyze the impact of inhibition of vesicle acidification on bacterial escape into the cytoplasm.
Background of Acidification of Microbial Vacuoles
A vesicle is an acidic compartment containing a wide variety of hydrolytic enzymes. Acidification within the vesicle plays a key role in many cellular processes, such as chemical osmotic regulation of cytoplasmic ion homeostasis and efficient sorting of vesicular proteins. The acidic environment within the lumen of the vesicle is maintained by the vesicle membrane H+- ATPase, where the intra-vesicular pH is an important signal for the activation of bacterial factors that mediate vesicle disruption and escape into the cytoplasm. In addition, it is critical that vesicles play a role in macromolecular degradation to maintain intracellular homeostasis. Autophagy, a major vesicle/lysosomal degradation system, is a vesicle/lysosomal protein hydrolysis process that is prevalent in many cell types.
Fig. 1. Acid tolerance mechanisms associated with cell membranes and ion transport systems. (Guan N, et al., 2020)
Acidification of Microbial Vacuoles Assessment Services
Vesicle pH is part of the genetic analysis of vesicle biogenesis and function in Saccharomyces cerevisiae. CD BioSciences focuses on measuring vesicle pH to identify mutants with defects in vesicle acidification. Our goal is to help you analyze the role of the ionic environment of phagosomal vesicles in the control of pathogens by macrophages.
To detect and analyze these mutants, our bioscientists develop customized processes for establishing vesicle pH assays using flow cytometry.
- Select bacteria with pH-sensitive genes and identify pH-driven promoters for these genes.
- Generate reporter strains that express green fluorescent protein (GFP) or other selected fluorescent proteins under a pH-responsive promoter.
- The strain is grown in a medium of known pH, generating a standard curve that converts the fluorescent signal to a specific pH.
Our Advantages
- We selected strains that could express different fluorescent proteins to facilitate quantitative analysis.
- The strains we select can be used to infect macrophages and provide feedback on environmental pH.
- Our services is widely used to develop studies on the manipulation of phagosome function by microorganisms.
- Our service can be used to explore resting and activated macrophage conditions as well as the phagosomal environment of bacteria.
We look forward to collaborating with you. You are always welcome to engage in discussions with us at any point of the project. If you are interested in our services, please feel free to contact us for more information.
Reference
- Guan N, Liu L. (2020) Microbial response to acid stress: mechanisms and applications. Appl Microbiol Biotechnol. 104(1):51-65.