Hai-Bin Luo, Aleksandra A Knapik, Janusz J Petkowski, Matthew Demas, Igor A Shumilin, Heping Zheng, Maksymilian Chruszcz, Wladek Minor
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The protein subunit consists of an N-terminal alpha-helical domain connected to a C-terminal LβH domain. CoA binds in the active site formed by the residues from adjacent LβH domains. After determination of CoA-bound structure, molecular dynamics simulations were performed to model the binding of AcCoA. Binding of both AcCoA and CoA to SACOL2570 was verified by isothermal titration calorimetry. SACOL2570 most likely acts as an acetyltransferase, using AcCoA as an acetyl group donor and an as-yet-undetermined chemical moiety as an acceptor. 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Here, we report the apo- and CoA-bound crystal structures of a member of the galactoside acetyltransferase superfamily from methicillin-resistant S. aureus SACOL2570 which was recently shown to be down regulated in S. aureus grown in the presence of fusidic acid, an antibiotic used to treat MRSA infections. SACOL2570 forms a homotrimer in solution, as confirmed by small-angle X-ray scattering and dynamic light scattering. The protein subunit consists of an N-terminal alpha-helical domain connected to a C-terminal LβH domain. CoA binds in the active site formed by the residues from adjacent LβH domains. After determination of CoA-bound structure, molecular dynamics simulations were performed to model the binding of AcCoA. Binding of both AcCoA and CoA to SACOL2570 was verified by isothermal titration calorimetry. SACOL2570 most likely acts as an acetyltransferase, using AcCoA as an acetyl group donor and an as-yet-undetermined chemical moiety as an acceptor. 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引用次数: 0
摘要
耐甲氧西林金黄色葡萄球菌(MRSA)是导致人类无数隐匿性和难治性感染的主要原因,尤其是在免疫系统受损的患者和儿童中。在这里,我们报告了耐甲氧西林金黄色葡萄球菌 SACOL2570 中半乳糖苷乙酰转移酶超家族成员的载脂蛋白和 CoA 结合晶体结构,最近的研究表明,在有治疗 MRSA 感染的抗生素--夫西地酸--存在的情况下,金黄色葡萄球菌会对 SACOL2570 进行下调。小角 X 射线散射和动态光散射证实,SACOL2570 在溶液中形成同源三聚体。蛋白质亚基由一个 N 端 alpha-helical 结构域和一个 C 端 LβH 结构域组成。CoA 与相邻 LβH 结构域残基形成的活性位点结合。在确定了 CoA 结合结构之后,对 AcCoA 的结合进行了分子动力学模拟。等温滴定量热法验证了 AcCoA 和 CoA 与 SACOL2570 的结合。SACOL2570 很可能是一种乙酰转移酶,以 AcCoA 作为乙酰基供体,以一种尚未确定的化学分子作为受体。SACOL2570 最近被用作突变的支架,导致产生笼状装配,并有可能用于产生更复杂的纳米结构。
Biophysical analysis of the putative acetyltransferase SACOL2570 from methicillin-resistant Staphylococcus aureus.
Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of a myriad of insidious and intractable infections in humans, especially in patients with compromised immune systems and children. Here, we report the apo- and CoA-bound crystal structures of a member of the galactoside acetyltransferase superfamily from methicillin-resistant S. aureus SACOL2570 which was recently shown to be down regulated in S. aureus grown in the presence of fusidic acid, an antibiotic used to treat MRSA infections. SACOL2570 forms a homotrimer in solution, as confirmed by small-angle X-ray scattering and dynamic light scattering. The protein subunit consists of an N-terminal alpha-helical domain connected to a C-terminal LβH domain. CoA binds in the active site formed by the residues from adjacent LβH domains. After determination of CoA-bound structure, molecular dynamics simulations were performed to model the binding of AcCoA. Binding of both AcCoA and CoA to SACOL2570 was verified by isothermal titration calorimetry. SACOL2570 most likely acts as an acetyltransferase, using AcCoA as an acetyl group donor and an as-yet-undetermined chemical moiety as an acceptor. SACOL2570 was recently used as a scaffold for mutations that lead the generation of cage-like assemblies, and has the potential to be used for the generation of more complex nanostructures.