S100 Proteins in the Innate Immune Response to Pathogens

Area of Science:

• Biochemistry
• Immunology
• Molecular Biology

Background:

• S100 proteins are calcium-binding proteins involved in various cellular processes.
• Specific S100 proteins (S100A7, S100A12, S100A8, S100A9) are critical in the innate immune response.
• These proteins employ nutritional immunity by depleting essential metals for pathogens and act as DAMPs activating immune receptors.

Purpose of the Study:

• To present protocols for the production and characterization of key S100 proteins involved in innate immunity.
• To enable high-level recombinant protein production.
• To facilitate the measurement of metal-binding affinities and antimicrobial activity.

Main Methods:

• High-level production of recombinant S100 proteins.
• Isothermal titration calorimetry (ITC) for measuring binding affinities to transition metals.
• Antimicrobial assays to evaluate the functional activity of S100 proteins.

Main Results:

• Established protocols for producing functional S100 proteins.
• Quantified the binding affinities of S100 proteins for transition metals.
• Demonstrated the antimicrobial activity of S100 proteins through functional assays.

Conclusions:

• The presented protocols are valuable for researchers studying S100 proteins in the context of innate immunity and nutritional immunity.
• These methods allow for detailed investigation of S100 protein function in pathogen defense.
• Further research can build upon these protocols to explore S100 protein roles in health and disease.