Nitrous oxide reductase from denitrifying Pseudomonas stutzeri. Genes for copper-processing and properties of the deduced products, including a new member of the family of ATP/GTP-binding proteins.
Nitrous oxide reductase from denitrifying Pseudomonas stutzeri. Genes for copper-processing and properties of the deduced products, including a new member of the family of ATP/GTP-binding proteins.
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DOI：
10.1111/j.1432-1033.1990.tb19265.x
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发表时间：
1990-09
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影响因子：
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通讯作者：
W. Zumft;Adelheid Viebrock-Sambale;C. Braun
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文献类型：
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作者：
W. Zumft;Adelheid Viebrock-Sambale;C. Braun
厌氧细菌施氏假单胞菌（Pseudomonasstutzeri）的一氧化二氮（N2 O）呼吸需要合成多铜酶N2 O还原酶，nosZ是该酶的结构基因，是携带几个必需基因的约8 kbp的DNA区域的一部分。插入突变的区域下游的nosZ产生脱辅基酶合成菌株，这就认为存在的功能，铜收购或铜加工，或两者兼而有之，N2 O还原酶。对约3.2 kbp的相关DNA区域进行测序，发现由三个基因组成，nosDFY，推测在单个转录单位内。细胞浓度，铜含量，和N2 O还原酶蛋白的位置进行了研究与突变体中的三个基因的影响。推导的基因产物之间的相互作用，并与细胞质膜似乎是类似的组件的抗生素休克敏感的细菌运输系统。nosF基因的33.8-kDa产物属于核苷酸结合蛋白家族[C. Higgins等人（1986）Nature 323，448-450]。其氨基酸序列显示两个典型的核苷酸结合基序，并且与该家族成员的氨基酸位置同一性约为30%。nosY基因的29.4-kDa产物是具有六个预测的跨膜螺旋和输出信号的疏水蛋白。nosD基因的48.2-kDa产物是周质组分;它携带输出信号并且是亲水性蛋白。N2 O还原酶本身是一种周质酶。我们的研究结果提供了一个辅助系统的铜加工的证据，并建议其参与周质生物合成的铜中心的N2 O还原酶。
Nitrous oxide (N2O) respiration by the denitrifying bacterium Pseudomonas stutzeri requires the synthesis of the multi-copper enzyme N2O reductase, nosZ, the structural gene for this enzyme, is part of a DNA region of approximately 8 kbp that carries several essential genes. Insertional mutagenesis of the region downstream of nosZ generates apoenzyme-synthesizing strains, which argues for the existence of functions for copper acquisition or copper processing, or both, for N2O reductase. The relevant DNA region of approximately 3.2 kbp was sequenced and found to consist of three genes, nosDFY, presumably within a single transcriptional unit. Cellular concentration, copper content, and location of the N2O reductase protein were studied with mutants which were affected in the three genes. Interactions of the deduced gene products among each other and with the cytoplasmic membrane appear to be analogous to those of the components of osmotic-shock-sensitive bacterial transport systems. The 33.8-kDa product of the nosF gene belongs to the family of nucleotide-binding proteins [C. Higgins et al. (1986) Nature 323, 448-450]. Its amino acid sequence shows two canonical nucleotide-binding motifs, and the positional identity of amino acids to members of this family is around 30%. The 29.4-kDa product of the nosY gene is a hydrophobic protein with six predicted transmembrane helices and an export signal. The 48.2-kDa product of the nosD gene is a periplasmic component; it carries an export signal and is a hydrophilic protein. N2O reductase itself is a periplasmic enzyme. Our results provide evidence for an auxiliary system of Cu processing and suggest its involvement in the periplasmic biosynthesis of the Cu centers of N2O reductase.