Crystallization and preliminary characterization of a novel haem-binding protein of Streptomyces reticuli.
Crystallization and preliminary characterization of a novel haem-binding protein of Streptomyces reticuli.
复制标题
网状链霉菌新型血红素结合蛋白的结晶和初步表征。
DOI：
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发表时间：
2008
期刊：
影响因子：
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通讯作者：
D. Ortiz de Orué Lucana
中科院分区：
文献类型：
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作者：
P. Zou;M. Groves;S. Viale;D. Ortiz de Orué Lucana
网状链霉菌是一种生长在土壤中的革兰氏阳性细菌，已被证明能分泌一种新的血红素结合蛋白，即HBPs。序列分析表明，HBPs的同源物存在于多种细菌中，包括不同的放线杆菌以及革兰氏阴性霍乱弧菌和肺炎克雷伯菌。当网状沙门氏菌在天然抗生素胡敏(Fe3+氧化形式的血红素)存在下培养时，体内HBPs的产量大大增加。突变分析表明，HBPs显著提高了网状沙门氏菌对毒性浓度胡敏素的抗性。先前的数据表明，新发现的双组分传感器系统SENS-SenR的存在也大大增强了网状沙门氏菌对Hamin和氧化还原循环化合物Pumbagin的抗性，这表明它在感知氧化还原变化中发挥了作用。HBPs和SENS-SenR之间的特异性相互作用在体外已经被证明，它调节过氧化氢酶-过氧化物酶CPEB以及HBPs的表达。HBPs在P2(1)3空间群中被重组过表达、纯化和结晶，晶胞边缘为152.5 A。晶体的衍射数据记录到最大分辨率为2.25A，并使用SAD方法从短暂浸泡在高浓度溴化钠中的晶体中获得了物相。
Streptomyces reticuli is a soil-growing Gram-positive bacteria that has been shown to secrete a novel haem-binding protein known as HbpS. Sequence analysis reveals that homologues of HbpS are found in a wide variety of bacteria, including different Actinobacteria and the Gram-negative Vibrio cholera and Klebsiella pneumoniae. The in vivo production of HbpS is greatly increased when S. reticuli is cultured in the presence of the natural antibiotic haemin (Fe3+ oxidized form of haem). Mutational analysis demonstrated that HbpS significantly increases the resistance of S. reticuli to toxic concentrations of haemin. Previous data show that the presence of the newly identified two-component sensor system SenS-SenR also considerably enhances the resistance of S. reticuli to haemin and the redox-cycling compound plumbagin, suggesting a role in the sensing of redox changes. Specific interaction between HbpS and SenS-SenR, which regulates the expression of the catalase-peroxidase CpeB, as well as HbpS, has been demonstrated in vitro. HbpS has been recombinantly overexpressed, purified and crystallized in space group P2(1)3, with a cell edge of 152.5 A. Diffraction data were recorded to a maximal resolution of 2.25 A and phases were obtained using the SAD method from crystals briefly soaked in high concentrations of sodium bromide.