Molecular evolution of the reactive oxygen-generating NADPH oxidase (Nox/Duox) family of enzymes.
Molecular evolution of the reactive oxygen-generating NADPH oxidase (Nox/Duox) family of enzymes.
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DOI：
10.1186/1471-2148-7-109
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发表时间：
2007-07-06
影响因子：
3.4
通讯作者：
Lambeth JD
中科院分区：
文献类型：
--
作者：
Kawahara T;Quinn MT;Lambeth JD
NADPH氧化酶（Nox）及其相关的双氧化酶（Duox）通过调节活性氧（ROS）的产生发挥多种生物学和病理学作用。Nox/Duox家族的成员已经在多种生物体中鉴定，包括哺乳动物、线虫、果蝇、绿色植物、真菌和黏菌;然而，对这些酶的分子进化历史知之甚少。我们收集并分析了来自脊椎动物、尾索动物、棘皮动物、昆虫、线虫、真菌、黏菌、阿米巴和植物等25个物种的101个Nox/Duox同源基因的推导氨基酸序列。与原核生物中存在的ROS防御酶（如超氧化物歧化酶和过氧化氢酶）相反，ROS生成的Nox/Duox直系同源物仅在进化后期出现。分子分类学揭示了Noxes和Duoxes的七个不同的亚科。钙调节的直系同源物代表4个亚科，分化较早，在生物学中分布最广。亚单位调节的Noxes代表了第二个主要的细分，并首次出现在真菌和变形虫。Nox 5在啮齿动物中丢失，而在内耳中起重力感知作用的Nox 3最近出现，对应于脊椎动物对陆地的全时适应。海胆Strongylocentrotus purpuratus拥有最早的Nox 2共同直系同源的脊椎动物Nox 1，2和3，而Nox 4首次出现稍晚于尾索动物。进化替换率的比较表明，Nox 2，调节亚基p47 phox和p67 phox，和Duox在脊椎动物中比其他Nox和Nox调节亚基更严格保守。氨基酸序列比较确定了关键的催化或调控区域，因为所有Nox/Duox直系同源物中有68个残基高度保守，其中14个与X连锁慢性肉芽肿病变体中Nox 2突变的残基相同。除了典型的基序，B-环，TM 6-FAD，VXGPFG基序，和极端的C-末端区域被确定为重要的Nox活性，通过突变分析验证。这些非典型的，但高度保守的区域的存在表明，所有的Nox/Duox可能具有共同的生物学功能，在Nox/Duox进化的漫长历史中仍然存在。该报告首次全面分析了Nox和Duox家族成员的进化和保守功能，包括保守氨基酸残基的鉴定。这些结果为今后的结构-功能研究和了解这些酶的生物学功能的演变提供了指导。
NADPH-oxidases (Nox) and the related Dual oxidases (Duox) play varied biological and pathological roles via regulated generation of reactive oxygen species (ROS). Members of the Nox/Duox family have been identified in a wide variety of organisms, including mammals, nematodes, fruit fly, green plants, fungi, and slime molds; however, little is known about the molecular evolutionary history of these enzymes. We assembled and analyzed the deduced amino acid sequences of 101 Nox/Duox orthologs from 25 species, including vertebrates, urochordates, echinoderms, insects, nematodes, fungi, slime mold amoeba, alga and plants. In contrast to ROS defense enzymes, such as superoxide dismutase and catalase that are present in prokaryotes, ROS-generating Nox/Duox orthologs only appeared later in evolution. Molecular taxonomy revealed seven distinct subfamilies of Noxes and Duoxes. The calcium-regulated orthologs representing 4 subfamilies diverged early and are the most widely distributed in biology. Subunit-regulated Noxes represent a second major subdivision, and appeared first in fungi and amoeba. Nox5 was lost in rodents, and Nox3, which functions in the inner ear in gravity perception, emerged the most recently, corresponding to full-time adaptation of vertebrates to land. The sea urchin Strongylocentrotus purpuratus possesses the earliest Nox2 co-ortholog of vertebrate Nox1, 2, and 3, while Nox4 first appeared somewhat later in urochordates. Comparison of evolutionary substitution rates demonstrates that Nox2, the regulatory subunits p47phox and p67phox, and Duox are more stringently conserved in vertebrates than other Noxes and Nox regulatory subunits. Amino acid sequence comparisons identified key catalytic or regulatory regions, as 68 residues were highly conserved among all Nox/Duox orthologs, and 14 of these were identical with those mutated in Nox2 in variants of X-linked chronic granulomatous disease. In addition to canonical motifs, the B-loop, TM6-FAD, VXGPFG-motif, and extreme C-terminal regions were identified as important for Nox activity, as verified by mutational analysis. The presence of these non-canonical, but highly conserved regions suggests that all Nox/Duox may possess a common biological function remained in a long history of Nox/Duox evolution. This report provides the first comprehensive analysis of the evolution and conserved functions of Nox and Duox family members, including identification of conserved amino acid residues. These results provide a guide for future structure-function studies and for understanding the evolution of biological functions of these enzymes.