Evaluation of the type I signal peptidase as antibacterial target for biofilm-associated infections of Staphylococcus epidermidis
Evaluation of the type I signal peptidase as antibacterial target for biofilm-associated infections of Staphylococcus epidermidis
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DOI：
10.1099/mic.0.031765-0
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发表时间：
2009-11-01
期刊：
影响因子：
2.8
通讯作者：
Van Aerschot, Arthur
中科院分区：
文献类型：
--
作者：
Bockstael, Katrijn;Geukens, Nick;Van Aerschot, Arthur
抗生素耐药性的发展是不可避免的，也是医院和社区的主要关注点。此外，生物膜生长的细菌对抗微生物处理不太敏感。在这方面，革兰氏阳性表皮葡萄球菌是医院生物膜相关感染的重要来源。在寻找新的抗菌疗法中，I型信号肽酶（SPase I）作为开发具有新作用模式的抗菌药物的潜在靶点。这种酶从分泌的蛋白质中切割出信号肽，使其对蛋白质分泌至关重要，因此对细菌细胞活力至关重要。S. epidermidis编码三种推定的SPase I（表示为Sip 1、Sip 2和Sip 3），其中Sip 1缺乏催化赖氨酸。本文对活性S. epidermidis SPases I更详细。Sip 2和Sip 3被发现补充温度敏感的大肠杆菌lepB突变体，证明其在体内的功能活性。使用基于荧光共振能量转移（FRET）的测定进一步说明了纯化的Sip 2和Sip 3蛋白的体外功能活性以及SPase I抑制剂芳基霉素A（2）对其活性的抑制。此外，我们证明了SPase I不仅是开发针对自由生活细菌的新型抗菌药物的有吸引力的靶标，而且是生物膜相关感染的可行靶标。
The development of antibacterial resistance is inevitable and is a major concern in hospitals and communities. Moreover, biofilm-grown bacteria are less sensitive to antimicrobial treatment. In this respect, the Gram-positive Staphylococcus epidermidis is an important source of nosocomial biofilm-associated infections. In the search for new antibacterial therapies, the type I signal peptidase (SPase I) serves as a potential target for development of antibacterials with a novel mode of action. This enzyme cleaves off the signal peptide from secreted proteins, making it essential for protein secretion, and hence for bacterial cell viability. S. epidermidis encodes three putative SPases I (denoted Sip1, Sip2 and Sip3), of which Sip1 lacks the catalytic lysine. In this report, we investigated the active S. epidermidis SPases I in more detail. Sip2 and Sip3 were found to complement a temperature-sensitive Escherichia coli lepB mutant, demonstrating their in vivo functional activity. In vitro functional activity of purified Sip2 and Sip3 proteins and inhibition of their activity by the SPase I inhibitor arylomycin A(2) were further illustrated using a fluorescence resonance energy transfer (FRET)-based assay. Furthermore, we demonstrated that SPase I not only is an attractive target for development of novel antibacterials against free-living bacteria, but also is a feasible target for biofilm-associated infections.