A new crystal structure of the bifunctional antibiotic simocyclinone D8 bound to DNA gyrase gives fresh insight into the mechanism of inhibition.
A new crystal structure of the bifunctional antibiotic simocyclinone D8 bound to DNA gyrase gives fresh insight into the mechanism of inhibition.
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DOI：
10.1016/j.jmb.2014.02.017
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发表时间：
2014-05-15
影响因子：
5.6
通讯作者：
Maxwell A
中科院分区：
文献类型：
--
作者：
Hearnshaw SJ;Edwards MJ;Stevenson CE;Lawson DM;Maxwell A
Simocyclinone D8（SD 8）是一种由链霉菌产生的抗生素，靶向DNA促旋酶。先前的SD 8结构与DNA促旋酶A蛋白（GyrA）的N端结构域复合，表明四个SD 8分子稳定了蛋白质的四聚体;随后的质谱实验表明，蛋白质二聚体与两个与DNA相关的SD 8更有可能。这项工作描述了进一步截短形式的GyrA N-末端结构域片段的结构，有和没有SD 8结合。具有SD 8的结构具有结合在相同GyrA二聚体内的两个SD 8分子。这种新的结构与GyrA中赋予SD 8抗性的突变完全一致，并且通过与GyrA N-末端结构域的新的载脂蛋白结构进行比较，揭示了SD 8结合后可能发生的构象变化以及SD 8抑制促旋酶的详细机制。等温滴定量热实验与晶体学结果一致，并进一步表明先前观察到的SD 8和GyrB之间的复合物比与GyrA的相互作用弱约1000倍。设计片段以揭示GyrA-药物复合物的生物学相关结构。这种结构充分解释了所有可用的生物化学/生物物理/遗传数据。GyrB中的结合位点比GyrA中的位点弱约1000倍。
Simocyclinone D8 (SD8) is an antibiotic produced by Streptomyces antibioticus that targets DNA gyrase. A previous structure of SD8 complexed with the N-terminal domain of the DNA gyrase A protein (GyrA) suggested that four SD8 molecules stabilized a tetramer of the protein; subsequent mass spectrometry experiments suggested that a protein dimer with two symmetry-related SD8s was more likely. This work describes the structures of a further truncated form of the GyrA N-terminal domain fragment with and without SD8 bound. The structure with SD8 has the two SD8 molecules bound within the same GyrA dimer. This new structure is entirely consistent with the mutations in GyrA that confer SD8 resistance and, by comparison with a new apo structure of the GyrA N-terminal domain, reveals the likely conformation changes that occur upon SD8 binding and the detailed mechanism of SD8 inhibition of gyrase. Isothermal titration calorimetry experiments are consistent with the crystallography results and further suggest that a previously observed complex between SD8 and GyrB is ~ 1000-fold weaker than the interaction with GyrA. Fragment engineered to reveal biologically relevant structure of GyrA–drug complex. This structure fully explains all available biochemical/biophysical/genetic data. Binding site in GyrB is ~ 1000-fold weaker than site in GyrA.
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影响因子：
5
作者：
Collin, Frederic;Karkare, Shantanu;Maxwell, Anthony
通讯作者：
Maxwell, Anthony
DOI：
10.1073/pnas.93.9.4057
发表时间：
1996-04-30
影响因子：
11.1
作者：
Roca, J;Berger, JM;Wang, JC
通讯作者：
Wang, JC
影响因子：
2.9
作者：
Edwards, Marcus J.;Williams, Mark A.;McKay, Adam R.
通讯作者：
McKay, Adam R.
影响因子：
2.9
作者：
ALI, JA;JACKSON, AP;MAXWELL, A
通讯作者：
MAXWELL, A
影响因子：
4.9
作者：
Sissi, C.;Vazquez, E.;Palumbo, M.
通讯作者：
Palumbo, M.