In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins.
In vitro reconstitution demonstrates the cell wall ligase activity of LCP proteins.
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DOI：
10.1038/nchembio.2302
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发表时间：
2017-04
影响因子：
14.8
通讯作者：
Walker S
中科院分区：
文献类型：
--
作者：
Schaefer K;Matano LM;Qiao Y;Kahne D;Walker S
球囊是一种肽聚糖基质，可保护细菌免受渗透溶解。在革兰氏阳性生物中，球囊被对生存重要的糖聚合物密集地功能化，但组装如何发生尚不清楚。在金黄色葡萄球菌中，三个LCP家族成员参与了将主要的糖聚合物壁磷壁酸（WTA）连接到肽聚糖上，但这些或任何其他LCP蛋白的连接酶活性尚未得到证实。使用WTA和肽聚糖底物产生chemoenzymatically，我们表明，所有这三种蛋白质可以转移WTA前体新生肽聚糖，建立LCP蛋白是肽聚糖glycopolymer连接酶。虽然所有的S。金黄色葡萄球菌LCP蛋白具有将WTA连接到PG的能力，我们表明它们的细胞功能不是冗余的。缺乏lcpA的菌株具有与WTA无效菌株相似的表型，表明这是最重要的WTA连接酶。本研究为进一步研究LCP酶参与细胞壁组装奠定了基础。
Sacculus is a peptidoglycan matrix that protects bacteria from osmotic lysis. In Gram-positive organisms, the sacculus is densely functionalized with glycopolymers important for survival, but how assembly occurs is not known. In Staphylococcus aureus, three LCP family members have been implicated in attaching the major glycopolymer wall teichoic acid (WTA) to peptidoglycan, but ligase activity has not been demonstrated for these or any other LCP proteins. Using WTA and peptidoglycan substrates produced chemoenzymatically, we show that all three proteins can transfer WTA precursors to nascent peptidoglycan, establishing that LCP proteins are peptidoglycan-glycopolymer ligases. Although all S. aureus LCP proteins have the capacity to attach WTA to PG, we show that their cellular functions are not redundant. Strains lacking lcpA have phenotypes similar to WTA null strains, indicating that this is the most important WTA ligase. This work provides a foundation for studying how LCP enzymes participate in cell wall assembly.