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哺乳动物中前所未有的 CYP74 酶的检测:蝙蝠中的过氧化物水解酶 CYP74C44 。

Detection of Unprecedented CYP74 Enzyme in Mammal: Hydroperoxide Lyase CYP74C44 of the Bat .

机构信息

Kazan Institute of Biochemistry and Biophysics, FRC Kazan Scientific Center of RAS, P.O. Box 30, 420111 Kazan, Russia.

出版信息

Int J Mol Sci. 2022 Jul 20;23(14):8009. doi: 10.3390/ijms23148009.

Abstract

The genome of the neotropical fruit bat was recently sequenced, revealing an unexpected gene encoding a plant-like protein, CYP74C44, which shares ca. 90% sequence identity with the putative CYP74C of . The preparation and properties of the recombinant CYP74C44 are described in the present work. The CYP74C44 enzyme was found to be active against the 13- and 9-hydroperoxides of linoleic and α-linolenic acids (13-HPOD, 13-HPOT, 9-HPOD, and 9-HPOT, respectively), as well as the 15-hydroperoxide of eicosapentaenoic acid (15-HPEPE). All substrates studied were specifically transformed into chain cleavage products that are typical for hydroperoxide lyases (HPLs). The HPL chain cleavage reaction was validated by the identification of NaBH-reduced products (Me/TMS) of 15-HPEPE and 13- and 9-hydroperoxides as (all-)-14-hydroxy-5,8,11-tetradecatrienoic, (9)-12-hydroxy-9-dodecenoic, and 9-hydroxynonanoic acids (Me/TMS), respectively. Thus, CYP74C44 possessed the HPL activity that is typical for the CYP74C subfamily proteins.

摘要

最近对新热带水果蝙蝠的基因组进行了测序,揭示了一个意想不到的基因,该基因编码一种类似植物的蛋白质 CYP74C44,其与 的推定 CYP74C 具有约 90%的序列同一性。本工作描述了重组 CYP74C44 的制备和性质。发现 CYP74C44 酶对亚油酸和α-亚麻酸的 13-和 9-过氧化物(分别为 13-HPOD、13-HPOT、9-HPOD 和 9-HPOT)以及二十碳五烯酸的 15-过氧化物(15-HPEPE)具有活性。研究的所有底物都特异性转化为典型的过氧化物裂解酶(HPL)的链裂解产物。通过鉴定 15-HPEPE 和 13-和 9-过氧化物的 NaBH 还原产物(Me/TMS)作为(all-)-14-羟基-5,8,11-十四碳三烯酸、(9)-12-羟基-9-十二烯酸和 9-羟基壬酸(Me/TMS),验证了 HPL 链裂解反应。因此,CYP74C44 具有 CYP74C 亚家族蛋白典型的 HPL 活性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/92b4/9320521/6e7331256496/ijms-23-08009-g001.jpg

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