Molecular dissection of phage lysin PlySs2: integrity of the catalytic and cell wall binding domains is essential for its broad lytic activity
Molecular dissection of phage lysin PlySs2: integrity of the catalytic and cell wall binding domains is essential for its broad lytic activity
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噬菌体溶素 PlySs2 的分子解剖：催化和细胞壁结合域的完整性对其广泛的裂解活性至关重要。
DOI：
10.1007/s12250-014-3535-6
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发表时间：
2015-02-01
影响因子：
5.5
通讯作者：
Wei, Hongping
中科院分区：
文献类型：
--
作者：
Huang, Yanling;Yang, Hang;Wei, Hongping
据报道，新型噬菌体溶素PlySs2对各种细菌具有高度活性，包括葡萄球菌、链球菌和李斯特菌。然而，其广泛的裂解谱的分子机制仍有待建立。在本研究中，裂解活性的催化结构域（CD，PlySc）和细胞壁结合结构域（CBD，PlySb）的PlySs2的结合特异性进行了检查。我们的结果表明，单独的PlySc保持非常有限的裂解活性。融合增强型绿色荧光蛋白（EGFP）的PlySb对所测试的链球菌菌株（包括S. suis，S. dysgalactiae和S.无乳链球菌，而不是葡萄球菌，支持其作为链球菌靶向溶素工程的良好CBD供体的效用。EGFP融合的完整PlyS2同样显示出对链球菌的高亲和力，但对葡萄球菌没有。值得注意的是，四个截短的PlySb片段显示没有结合能力。这些发现共同表明PlySc和PlySb结构域的完整性是PlySs2的广泛裂解活性的重要决定因素。
The novel phage lysin PlySs2, is reported to be highly active against various bacteria, including staphylococci, streptococci and Listeria. However, the molecular mechanisms underlying its broad lytic spectrum remain to be established. In the present study, the lytic activity of the catalytic domain (CD, PlySc) and binding specificity of the cell wall binding domain (CBD, PlySb) of PlySs2 were examined. Our results showed that PlySc alone maintains very limited lytic activity. Enhanced green fluorescent protein (EGFP)-fused PlySb displayed high binding affinity to the streptococcal strains tested, including S. suis, S. dysgalactiae, and S. agalactiae, but not staphylococci, supporting its utility as a good CBD donor for streptococcal-targeted lysin engineering. EGFP-fused intact PlySs2 similarly displayed high affinity for streptococci, but not staphylococci. Notably, four truncated PlySb fragments showed no binding capacity. These findings collectively indicate that integrity of the PlySc and PlySb domains is an essential determinant of the broad lytic activity of PlySs2.