Application of a bacterial two-hybrid system for the analysis of protein-protein interactions between FemABX family proteins
Application of a bacterial two-hybrid system for the analysis of protein-protein interactions between FemABX family proteins
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DOI：
10.1099/mic.0.26315-0
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发表时间：
2003-10-01
期刊：
影响因子：
2.8
通讯作者：
Berger-Bächi, B
中科院分区：
文献类型：
--
作者：
Rohrer, S;Berger-Bächi, B
蛋白质之间的相互作用在所有细胞过程中都扮演着重要的角色。酵母和细菌中双杂交系统的发展使得在体内评估这种相互作用成为可能。利用最近开发的细菌双杂交系统，分析了FemABX蛋白家族成员金黄色葡萄球菌蛋白FemA、FemB和FmhB之间的相互作用，FemABX蛋白参与了许多革兰氏阳性菌的肽聚糖生物合成和β-内酰胺耐药。FmhB参与金黄色葡萄球菌肽聚糖五甘氨酸间肽的甘氨酸1的加成，FEMA和FemB分别对2/3和4/5的甘氨酸具有专一性。FEMA-FEMA、FEMA-FemB和FemB-FemB相互作用，而FmHB仅以单体形式存在。通过谷胱甘肽S转移酶下拉试验和凝胶过滤证实了细菌双杂交系统检测到的相互作用。
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.