Actinobacillus utilizes a binding protein-dependent ABC transporter to acquire the active form of vitamin B(6).
Actinobacillus utilizes a binding protein-dependent ABC transporter to acquire the active form of vitamin B(6).
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DOI：
10.1016/j.jbc.2021.101046
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发表时间：
2021-09
期刊：
影响因子：
--
通讯作者：
Moraes TF
中科院分区：
文献类型：
--
作者：
Pan C;Zimmer A;Shah M;Huynh MS;Lai CC;Sit B;Hooda Y;Curran DM;Moraes TF
细菌需要高效率的吸收系统，以在营养有限的环境中生存和增殖，例如在宿主生物体中发现的那些。细菌质膜中的ABC转运蛋白提供了许多底物的转运机制。在这项研究中，我们研究了一个操纵子包含一个周质结合蛋白在放线杆菌的营养收购的潜在作用。从周质结合蛋白的晶体结构获得的1.76 μ m分辨率的电子密度图与蛋白结合位点内含有吡哆醛-5 ′-磷酸（P5 P/吡哆醛磷酸/维生素B6的活性形式）配体的分子模型最佳拟合。通过等温滴定量热法、微量热泳和质谱法验证了与该周质结合蛋白结合的P5 P的身份，从而使我们命名蛋白质P5 PA和操纵子P5 PAB。为了说明这个摄取系统的功能效用，我们引入了P5 PAB操纵子从放线杆菌胸膜肺炎到大肠杆菌K-12菌株，这是缺乏P5 P合成所需的关键酶。该菌株在低水平P5 P下的生长支持该操纵子在P5 P摄取中的功能作用。这是第一次报道一个专门的P5 P细菌摄取系统，但通过生物信息学，我们发现同源物主要是在致病性代表巴斯德菌科，这表明这种操纵子更广泛地存在于放线杆菌属以外。
Bacteria require high-efficiency uptake systems to survive and proliferate in nutrient-limiting environments, such as those found in host organisms. ABC transporters in the bacterial plasma membrane provide a mechanism for transport of many substrates. In this study, we examine an operon containing a periplasmic binding protein in Actinobacillus for its potential role in nutrient acquisition. The electron density map of 1.76 Å resolution obtained from the crystal structure of the periplasmic binding protein was best fit with a molecular model containing a pyridoxal-5′-phosphate (P5P/pyridoxal phosphate/the active form of vitamin B6) ligand within the protein's binding site. The identity of the P5P bound to this periplasmic binding protein was verified by isothermal titration calorimetry, microscale thermophoresis, and mass spectrometry, leading us to name the protein P5PA and the operon P5PAB. To illustrate the functional utility of this uptake system, we introduced the P5PAB operon from Actinobacillus pleuropneumoniae into an Escherichia coli K-12 strain that was devoid of a key enzyme required for P5P synthesis. The growth of this strain at low levels of P5P supports the functional role of this operon in P5P uptake. This is the first report of a dedicated P5P bacterial uptake system, but through bioinformatics, we discovered homologs mainly within pathogenic representatives of the Pasteurellaceae family, suggesting that this operon exists more widely outside the Actinobacillus genus.
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影响因子：
9.4
作者：
Gautier, AL;Dubois, D;Gaillot, O
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Gaillot, O
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