Stabilization of the primary sigma factor of Staphylococcus aureus by core RNA polymerase.

Stabilization of the primary sigma factor of Staphylococcus aureus by core RNA polymerase.
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DOI:
10.5483/bmbrep.2010.43.3.176
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发表时间:
2010-03
期刊:
影响因子:
3.8
通讯作者:
Sau S
Sau S
中科院分区:
生物学3区
文献类型:
--
作者:
Mondal R;Ganguly T;Chanda PK;Bandhu A;Jana B;Sau K;Lee CY;Sau S

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金黄色葡萄球菌的主要西格玛因子(σA)是一个潜在的药物靶点,但在结构水平上很少被研究。使用 N 末端组氨酸标记的 σA (His-σA),我们证明它在溶液中以单体形式存在,拥有多个结构域,主要包含 α 螺旋,并在核心 RNA 聚合酶存在的情况下有效结合金黄色葡萄球菌启动子 DNA。虽然 HisσA 的 N 端和 C 端本质上都是柔性的,但其 DNA 结合区域中的两个 Trp 残基被掩埋。将孵育温度从 25°C 升高至 40°C 后,约 60% 的输入 His-σA 被嗜热菌蛋白酶裂解。 His-σA 的聚集也在 45oC 时迅速开始。根据平衡展开实验,His-σA 稳定的吉布斯自由能估计为+0.70 kcal mol-1。这些数据共同表明金黄色葡萄球菌的初级西格玛因子是一种不稳定的蛋白质。然而,核心RNA聚合酶明显稳定了σA。 [BMB 2010 年报告; 43(3):176–181]
The primary sigma factor (σA) of Staphylococcus aureus, a potential drug target, was little investigated at the structural level. Using an N-terminal histidine-tagged σA (His-σA), here we have demonstrated that it exits as a monomer in solution, possesses multiple domains, harbors primarily α-helix and efficiently binds to a S. aureus promoter DNA in the presence of core RNA polymerase. While both Nand C-terminal ends of HisσA are flexible in nature, two Trp residues in its DNA binding region are buried. Upon increasing the incubation temperature from 25° to 40°C, ~60% of the input His-σA was cleaved by thermolysin. Aggregation of His-σA was also initiated rapidly at 45oC. From the equilibrium unfolding experiment, the Gibbs free energy of stabilization of His-σA was estimated to be +0.70 kcal mol-1. The data together suggest that primary sigma factor of S. aureus is an unstable protein. Core RNA polymerase however stabilized σA appreciably. [BMB reports 2010; 43(3): 176–181]
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