洞察果蝇CYP4G酶的独特特征。

IF 3.2 2区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Insect Biochemistry and Molecular Biology Pub Date : 2023-11-25 DOI:10.1016/j.ibmb.2023.104041
Mary Kefi , Parasyris Konstantinos , Vasileia Balabanidou , Chara Sarafoglou , Dimitra Tsakireli , Vassilis Douris , Maria Monastirioti , Jean-Didier Maréchal , René Feyereisen , John Vontas
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引用次数: 0

摘要

CYP4G亚家族的细胞色素P450酶在结构和功能方面是一些最有趣的昆虫P450酶。在果蝇中,CYP4G1在卵泡细胞中高表达,是角质层碳氢化合物生物合成的最后一种酶,而CYP4G15在大脑中表达,功能未知。这两种蛋白都有一个cyp4g特异性和特征氨基酸序列插入,对应于G和H螺旋之间的环,其功能尚不清楚。在这里,我们研究了果蝇CYP4Gs的这些神秘的结构和功能特征。首先,我们使用反向遗传学方法产生了全部或部分从CYP4G1中去除cyp4g特异性环的D. melanogaster菌株。我们发现,P450的正常折叠并不需要完整的环,但它对功能至关重要,而且这种酶制造碳氢化合物的能力只需要6个氨基酸的一小段。其次,我们通过免疫细胞化学证实了CYP4G15在大脑中表达,并表明它与皮层胶质细胞亚型特异性相关。然后,我们在卵泡细胞中异位表达CYP4G15,揭示它可以产生碳氢化合物的混合物,尽管在数量上较低的水平导致CYP4G1敲低的苍蝇只能部分拯救。本文研究的CYP4G1结构变异有助于CYP4G酶的生化表征。我们的结果也提出了碳氢化合物的假定作用和他们的合成皮层胶质细胞的问题。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Insights into unique features of Drosophila CYP4G enzymes

The cytochrome P450 enzymes of the CYP4G subfamily are some of the most intriguing insect P450s in terms of structure and function. In Drosophila, CYP4G1 is highly expressed in the oenocytes and is the last enzyme in the biosynthesis of cuticular hydrocarbons, while CYP4G15 is expressed in the brain and is of unknown function. Both proteins have a CYP4G-specific and characteristic amino acid sequence insertion corresponding to a loop between the G and H helices whose function is unclear. Here we address these enigmatic structural and functional features of Drosophila CYP4Gs. First, we used reverse genetics to generate D. melanogaster strains in which all or part of the CYP4G-specific loop was removed from CYP4G1. We showed that the full loop was not needed for proper folding of the P450, but it is essential for function, and that just a short stretch of six amino acids is required for the enzyme’s ability to make hydrocarbons. Second, we confirmed by immunocytochemistry that CYP4G15 is expressed in the brain and showed that it is specifically associated with the cortex glia cell subtype. We then expressed CYP4G15 ectopically in oenocytes, revealing that it can produce of a blend of hydrocarbons, albeit to quantitatively lower levels resulting in only a partial rescue of CYP4G1 knockdown flies. The CYP4G1 structural variants studied here should facilitate the biochemical characterization of CYP4G enzymes. Our results also raise the question of the putative role of hydrocarbons and their synthesis by cortex glial cells.

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来源期刊
CiteScore
7.40
自引率
5.30%
发文量
105
审稿时长
40 days
期刊介绍: This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.
期刊最新文献
Identification of a gene promoter active in Lucilia sericata larval salivary glands using a rapid transient expression assay. In vivo RNAi screening identifies multiple deubiquitinases required for the maintenance of intestinal homeostasis in Drosophila. JAK and STAT5B mediate olfactory response of migratory locusts to their own volatiles. Functional redundancy of the three insulin receptors of cockroaches. Editorial Board
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