Dual Mode of Action for Plusbacin A3 in Staphylococcus aureus.
Dual Mode of Action for Plusbacin A3 in Staphylococcus aureus.
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Plusbacin A3 对金黄色葡萄球菌的双重作用模式。
DOI：
10.1021/acs.jpcb.6b11039
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发表时间：
2017
期刊：
影响因子：
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通讯作者：
Schaefer,Jacob
中科院分区：
文献类型：
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作者：
O'Connor,RobertD;Singh,Manmilan;Chang,James;Kim,SungJoon;VanNieuwenhze,Michael;Schaefer,Jacob
我们使用C{F}， N{F}和N{P}旋转回声双共振NMR来确定19f和15n双标记plusbacin A3和双标记deslipop -plusbacin A3的位置和构象，它们分别与金黄色葡萄球菌在含有[1-13C]甘氨酸的培养基中生长的整个细胞的细胞壁结合。31p主要存在于壁苔酸中。大约25%的plusbacin头基（环状沉积肽主链）处于封闭构象（N-F分离为6 Å），而75%处于更开放的构象（N-F分离为12 Å）。封闭的头基与壁壁壁酸没有接触，而开放的头基与壁壁酸有很强的接触。这使得封闭的头团位于细胞壁的疏水区域，而开放的头团位于亲水区域。plusbacin尾部与壁壁壁酸或细胞膜的31p均无接触，因此位于细胞壁疏水区域。此外，plusbacin a3的头部和尾部都与细胞壁肽聚糖五酰桥接中的甘氨酸13c接触，并与膜表面附近的13c标记嘌呤接触。我们根据plusbacin A3的双重作用模式来解释这些结果：首先，封闭构象的plusbacin A3破坏最靠近膜表面的肽聚糖层，导致通过转糖基化抑制链延伸；其次，开放构象的plusbacin A3使细胞膜变薄和破坏（可能包括破坏嵌入膜中的ATP结合盒转运体），从而导致ATP释放到细胞壁的亲水区域，随后被plusbacin A3结合。
We have used C{F}, N{F}, and N{P} rotational-echo double resonance NMR to determine the location and conformation of19F and15N double-labeled plusbacin A3and of double-labeleddeslipo-plusbacin A3, each bound to the cell walls of whole cells ofStaphyloccocus aureusgrown in media containing [1-13C]glycine. The31P is primarily in wall teichoic acid. Approximately 25% of plusbacin headgroups (the cyclic depsipeptide backbone) are in a closed conformation (N–F separation of 6 Å), while 75% are in a more open conformation (N–F separation of 12 Å). The closed headgroups have no contact with wall teichoic acid, whereas the open headgroups have a strong contact. This places the closed headgroups in hydrophobic regions of the cell wall and the open headgroups in hydrophilic regions. None of the plusbacin tails have contact with the31P of either wall teichoic acid or the cell membrane and thus are in hydrophobic regions of the cell wall. In addition, both heads and tails of plusbacin A3have contact with the glycyl13C incorporated in cell-wall peptidoglycan pentaglycyl bridgesandwith13C-labeled purines near the membrane surface. We interpret these results in terms of a dual mode of action for plusbacin A3:first, disruption of the peptidoglycan layer nearest to the membrane surface by closed-conformation plusbacin A3leading to an inhibition of chain extension by transglycosylation;second, thinning and disruption of the membrane (possibly including disruption of ATP-binding cassette transporters embedded in the membrane) by open-conformation plusbacin A3, thereby leading to release of ATP to the hydrophilic regions of the cell wall and subsequent binding by plusbacin A3.