Profiling the Heme-Binding Proteomes of Bacteria Using Chemical Proteomics

Area of Science:

• Biochemistry
• Chemical Biology
• Microbiology

Background:

• Heme is an essential cofactor with diverse roles, including signaling and gene regulation.
• Heme homeostasis is critical for bacterial survival, making heme-binding proteins potential drug targets.
• The heme-binding proteome in bacteria remains largely uncharacterized beyond model organisms.

Purpose of the Study:

• To develop and implement a novel chemical proteomics method for global profiling of heme-binding proteins in live bacterial cells.
• To characterize the heme-binding proteome in both Gram-positive and Gram-negative bacteria.
• To provide a versatile tool for advancing research in bacterial heme biology.

Main Methods:

• Utilized a panel of clickable and photoaffinity heme-based probes.
• Applied the method for global profiling of heme-binding proteins in live bacterial cells.
• Demonstrated the method's applicability across different bacterial cell types.

Main Results:

• Successfully profiled 32-54% of the known heme-binding proteomes in Gram-positive and Gram-negative bacteria.
• Established a first-of-its-kind chemical proteomics strategy for identifying heme-binding proteins in live cells.
• Validated the method's simplicity and interchangeability across different bacterial systems.

Conclusions:

• The developed chemical proteomics strategy enables comprehensive profiling of bacterial heme-binding proteins.
• This approach significantly enhances the understanding of heme biology in bacteria.
• The method offers a valuable tool for identifying novel antibacterial drug targets by focusing on heme homeostasis.