The cell wall binding domain of Listeria bacteriophage endolysin PlyP35 recognizes terminal GlcNAc residues in cell wall teichoic acid
The cell wall binding domain of Listeria bacteriophage endolysin PlyP35 recognizes terminal GlcNAc residues in cell wall teichoic acid
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DOI：
10.1111/j.1365-2958.2011.07774.x
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发表时间：
2011-09-01
影响因子：
3.6
通讯作者：
Loessner, Martin J.
中科院分区：
文献类型：
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作者：
Eugster, Marcel R.;Haug, Martina C.;Loessner, Martin J.
噬菌体内溶素的细胞壁结合结构域（CBD）以非凡的特异性将酶靶向细菌肽聚糖中的其底物。尽管对这些酶作为新型抗菌剂的强烈兴趣，但关于它们与细菌壁及其结合配体的相互作用知之甚少。我们研究了李斯特菌噬菌体内溶素PlyP 35与肽聚糖上磷壁酸聚合物中存在的碳水化合物残基的相互作用。生物化学和遗传分析表明，CBD的PlyP 35特异性识别N-乙酰葡糖胺（GlcNAc）残基在位置C4的多核糖醇磷酸亚基。CBDP 35的结合可以通过从细胞壁去除壁磷壁酸（WTA）聚合物来防止，并且通过添加纯化的WTA或乙酰化的NTA来抑制。我们表明，单核细胞增生李斯特菌lmo 2549和lmo 2550所需的装饰WTA与GlcNAc。任一基因的失活导致缺乏GlcNAc糖基化，并且突变体未能结合CBDP 35。我们还报道了L.单核细胞增多症菌株WSLC 1442的突变是由于lmo 2550中的小缺失，导致合成了负责糖基化缺陷的截短基因产物。与lmo 2550互补完全恢复了特征性血清型1/2特异性WTA和野生型表型的展示。
The cell wall binding domains (CBD) of bacteriophage endolysins target the enzymes to their substrate in the bacterial peptidoglycan with extraordinary specificity. Despite strong interest in these enzymes as novel antimicrobials, little is known regarding their interaction with the bacterial wall and their binding ligands. We investigated the interaction of Listeria phage endolysin PlyP35 with carbohydrate residues present in the teichoic acid polymers on the peptidoglycan. Biochemical and genetic analyses revealed that CBD of PlyP35 specifically recognizes the N-acetylglucosamine (GlcNAc) residue at position C4 of the polyribitol-phosphate subunits. Binding of CBDP35 could be prevented by removal of wall teichoic acid (WTA) polymers from cell walls, and inhibited by addition of purified WTAs or acetylated saccharides. We show that Listeria monocytogenes genes lmo2549 and lmo2550 are required for decoration of WTAs with GlcNAc. Inactivation of either gene resulted in a lack of GlcNAc glycosylation, and the mutants failed to bind CBDP35. We also report that the GlcNAc-deficient phenotype of L. monocytogenes strain WSLC 1442 is due to a small deletion in lmo2550, resulting in synthesis of a truncated gene product responsible for the glycosylation defect. Complementation with lmo2550 completely restored display of characteristic serovar 1/2 specific WTA and the wild-type phenotype.