The toolbox of Auricularia auricula-judae dye-decolorizing peroxidase - Identification of three new potential substrate-interaction sites.

The toolbox of Auricularia auricula-judae dye-decolorizing peroxidase - Identification of three new potential substrate-interaction sites.
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黑木耳染料脱色过氧化物酶的工具箱 - 三个新的潜在底物相互作用位点的鉴定

DOI:
10.1016/j.abb.2014.12.016
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发表时间:
2015
影响因子:
3.9
通讯作者:
Plattner
Plattner
中科院分区:
生物学3区
文献类型:
--
作者:
Strittmatter;Serrer;Ullrich;Hofrichter;Piontek;Schleicher;Plattner

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染料脱色过氧化物酶(DyPs)如AauDyPI能氧化不同种类和大小的底物。anAauDyPI-咪唑复合物的晶体结构提供了深入了解有机分子的束缚模式内的血红素腔的DyP。几个小的含氮杂环芳香族化合物被证明结合在theAauDyPI血红素腔,暗示的DyPs的敏感性唑类抑制剂类似于细胞色素P450。咪唑被证实是一种竞争性抑制剂方面的过氧化物结合。相比之下,庞大的底物如蒽醌染料在酶表面转化。在晶体结构中,底物类似物4-(2-羟乙基)-1-哌嗪乙磺酸(HEPES)与富含酪氨酸的中空区结合,其中含有Y25、Y147和Y337。自旋捕获与一氧化氮供体揭示Y229作为一个额外的酪氨酸为基础的自由基中心inAauDyPI。多频EPR光谱进一步揭示了活化AauDyPI中存在至少一个中间体C377自由基中心,其中W377是最可能的候选者。
Dye-decolorizing peroxidases (DyPs) such asAauDyPI from the fungusAuricularia auricula-judaeare able to oxidize substrates of different kinds and sizes. A crystal structure of anAauDyPI–imidazole complex gives insight into the binding patterns of organic molecules within the heme cavity of a DyP. Several smallN-containing heterocyclic aromatics are shown to bind in theAauDyPI heme cavity, hinting to susceptibility of DyPs to azole-based inhibitors similar to cytochromes P450. Imidazole is confirmed as a competitive inhibitor with regard to peroxide binding. In contrast, bulky substrates such as anthraquinone dyes are converted at the enzyme surface. In the crystal structure a substrate analog, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), binds to a tyrosine-rich hollow harboring Y25, Y147, and Y337. Spin trapping with a nitric oxide donor uncovers Y229 as an additional tyrosine-based radical center inAauDyPI. Multi-frequency EPR spectroscopy further reveals the presence of at least one intermediate tryptophanyl radical center in activatedAauDyPI with W377 as the most likely candidate.
染料脱色过氧化物酶 DyP 的催化机制可能需要天冬氨酸残基的摆动运动
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