Leishmania major possesses a unique HemG-type protoporphyrinogen IX oxidase

Leishmania major possesses a unique HemG-type protoporphyrinogen IX oxidase
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大型利什曼原虫拥有独特的 HemG 型原卟啉原 IX 氧化酶

DOI:
10.1042/bsr20140081
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发表时间:
2014
期刊:
影响因子:
4
通讯作者:
Karrie
Karrie
中科院分区:
生物学3区
文献类型:
--
作者:
Zwerschke;Karrie

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主要利什曼原虫既可以利用宿主血红素,也可以利用宿主提供的前体部分合成四吡咯。然而,只有间接证据可用于这部分晚期血红素生物合成途径。在这里,我们证明了LMJF_06_1280基因的L.主要编码HemG型PPO(原卟啉原IX氧化酶),催化原卟啉原IX氧化为原卟啉IX。有趣的是,锥虫是目前唯一已知的具有HemG型酶的真核生物。LMJF_06_1280基因与LMJF_06_1270和LMJF_06_1290分别形成一个潜在的转录单位,前者编码粪卟啉原Ⅲ氧化酶,后者编码细胞色素5。大肠杆菌Δ hemG菌株LG 285的功能互补证实了hemG主基因的存在。在E.用HPLC分析观察到大肠杆菌。纯化的重组L. majorHemG在体外以不同的泛醌和三苯基四氮唑为电子受体显示PPO活性。FMN被鉴定为L.主要HemG辅因子。活性位点残基被发现是必不可少的HemG催化。这些数据与L. majorCPO和铁螯合酶活性的生理学证据为L.少校
Leishmania majorwas proposed to either utilize haem from its host or partially synthesize the tetrapyrrole from host provided precursors. However, only indirect evidence was available for this partial late haem biosynthetic pathway. Here, we demonstrate that the LMJF_06_1280 gene ofL. majorencodes a HemG-type PPO (protoporphyrinogen IX oxidase) catalysing the oxidation of protoporphyrinogen IX to protoporphyrin IX. Interestingly, trypanosomatids are currently the only known eukaryotes possessing HemG-type enzymes. The LMJF_06_1280 gene forms a potential transcriptional unit with LMJF_06_1270 encoding CPO (coproporphyrinogen III oxidase) and with LMJF_06_1290 for a cytochromeb5.In vivofunction of theL. major hemGgene was shown by the functional complementation of theEscherichia coli ΔhemGstrain LG285. Restored haem formation inE. coliwas observed using HPLC analyses. Purified recombinantL. majorHemG revealed PPO activityin vitrousing different ubiquinones and triphenyltetrazolium as electron acceptors. FMN was identified as theL. majorHemG cofactor. Active site residues were found to be essential for HemG catalysis. These data in combination with the solved crystal structures ofL. majorCPO and the physiological proof of a ferrochelatase activity provide clear-cut evidence for a partial haem biosynthetic pathway inL. major.
DOI: 10.4061/2011/873230
发表时间: 2011
期刊: Enzyme research
影响因子: --
作者:
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