Compartment-Specific Labeling of Bacterial Periplasmic Proteins by Peroxidase-Mediated Biotinylation
Open accessAbstract
The study of the bacterial periplasm requires techniques with sufficient spatial resolution and sensitivity to resolve the components and processes within this subcellular compartment. Peroxidase-mediated biotinylation has enabled targeted labeling of proteins within subcellular compartments of mammalian cells. We investigated whether this methodology could be applied to the bacterial periplasm. In this study, we demonstrated that peroxidase-mediated biotinylation can be performed in mycobacteria and Escherichia coli. To eliminate detection artifacts from natively biotinylated mycobacterial proteins, we validated two alternative labeling substrates, tyramide azide and tyramide alkyne, which enable biotin-independent detection of labeled proteins. We also targeted peroxidase expression to the periplasm, resulting in compartment-specific labeling of periplasmic versus cytoplasmic proteins in mycobacteria. Finally, we showed that this method can be used to validate protein relocalization to the cytoplasm upon removal of a secretion signal. This novel application of peroxidase-mediated protein labeling will advance efforts to characterize the role of the periplasm in bacterial physiology and pathogenesis.
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Identifying protein-protein interactions can provide insight on protein function.
“Identifying protein-protein interactions can provide enormous insight on protein function, as demonstrated by new in vivo proximity-labeling technologies.”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [161]
The paper describes live-cell proximity labeling and says that identifying periplasmic proteins may help elucidate the role of the periplasm (Abstract; page not supplied). The passages retrieved for “protein-protein interactions” and “protein function” address protein localization, not whether identifying interactions provides insight into a protein’s function. (LLM, conf=0.72)
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Self-citation
- Authors
- Ganapathy, U. S., Bai, L., Wei, L., Eckartt, K. A., Lett, C. M., Previti, M. L., Carrico, I. S., & Seeliger, J. C.
- Affiliations
- Stony Brook University (US)
- Corresponding author
- Seeliger, Jessica C.
- Venue
- ACS Infectious Diseases
- Publisher
- American Chemical Society (ACS)
- Published
- 2018-04-30
- Volume
- 4
- Issue
- 6
- Pages
- 918-925
- ISSN
- 2373-8227 (ISSN-L)
- PMC
- PMC6767932
- OpenAlex
- W2800962469
- Cited by
- 29
- References
- 35
- Citations by year
- 2026: 7 · 2025: 4 · 2024: 1 · 2023: 9 · 2022: 3 · 2021: 2 · 2019: 3
- FWCI
- 0.69
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- ACS Infectious Diseases (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of Allergy and Infectious Diseases (R21 AI126044)
- National Institute of Biomedical Imaging and Bioengineering (R21 EB01795501)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Biotin and Related Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Keywords
- periplasmic proteins, mycobacteria, Escherichia coli, protein relocalization, secretion signal, periplasm
- MeSH terms
- Bacterial Proteins, Biotin, Cytoplasm, Escherichia coli, Peroxidase, Recombinant Fusion Proteins, Staining and Labeling, Biotinylation, Periplasmic Proteins, Click Chemistry
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