Application of a bacterial two-hybrid system for the analysis of protein–protein interactions between FemABX family proteins | Zendy

Susanne Rohrer | Zendy; Brigitte BergerBächi | Zendy

Open Access

Application of a bacterial two-hybrid system for the analysis of protein–protein interactions between FemABX family proteins

Author(s) -

Susanne Rohrer,

Brigitte BergerBächi

Publication year - 2003

Publication title -

microbiology

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.352

H-Index - 35

eISSN - 1465-2080

pISSN - 1350-0872

DOI - 10.1099/mic.0.26315-0

Subject(s) - peptidoglycan , biochemistry , protein–protein interaction , two hybrid screening , bacteria , yeast , bacterial cell structure , staphylococcus aureus , biology , protein family , microbiology and biotechnology , chemistry , gene , genetics

Protein-protein interactions play an important role in all cellular processes. The development of two-hybrid systems in yeast and bacteria allows for in vivo assessment of such interactions. Using a recently developed bacterial two-hybrid system, the interactions of the Staphylococcus aureus proteins FemA, FemB and FmhB, members of the FemABX protein family, which is involved in peptidoglycan biosynthesis and beta-lactam resistance of numerous Gram-positive bacteria, were analysed. While FmhB is involved in the addition of glycine 1 of the pentaglycine interpeptide of S. aureus peptidoglycan, FemA and FemB are specific for glycines 2/3 and 4/5, respectively. FemA-FemA, FemA-FemB and FemB-FemB interactions were found, while FmhB exists solely as a monomer. Interactions detected by the bacterial two-hybrid system were confirmed using the glutathione S-transferase-pulldown assay and gel filtration.

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