Application of a bacterial two-hybrid system for the analysis of protein-protein interactions between FemABX family proteins.
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.
Rohrer
Susanne
S
University of Zürich, Institute of Medical Microbiology, Gloriastr. 32, CH-8028 Zürich, Switzerland.
Berger-Bächi
Brigitte
B
University of Zürich, Institute of Medical Microbiology, Gloriastr. 32, CH-8028 Zürich, Switzerland.
D001426:Bacterial Proteins / Q000737:chemistry* / Q000502:physiology; D002850:Chromatography, Gel; D019281:Dimerization; D008970:Molecular Weight; D013210:Staphylococcus / Q000737:chemistry*; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; C096686:FemA protein, Bacteria; C102928:FemB protein, Staphylococcus epidermidis