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Rational Redesign of Monoamine Oxidase A into a Dehydrogenase to Probe ROS in Cardiac Aging.
ACS Chemical Biology ( IF 4 ) Pub Date : 2020-06-26 , DOI: 10.1021/acschembio.0c00366
Luca Giacinto Iacovino 1 , Nicola Manzella 2 , Jessica Resta 2 , Maria Antonietta Vanoni 3 , Laura Rotilio 1 , Leonardo Pisani 4 , Dale Edward Edmondson 5 , Angelo Parini 2 , Andrea Mattevi 1 , Jeanne Mialet-Perez 2 , Claudia Binda 1
Affiliation  

Cardiac senescence is a typical chronic frailty condition in the elderly population, and cellular aging is often associated with oxidative stress. The mitochondrial-membrane flavoenzyme monoamine oxidase A (MAO A) catalyzes the oxidative deamination of neurotransmitters, and its expression increases in aged hearts. We produced recombinant human MAO A variants at Lys305 that play a key role in O2 reactivity leading to H2O2 production. The K305Q variant is as active as the wild-type enzyme, whereas K305M and K305S have 200-fold and 100-fold lower kcat values and similar Km. Under anaerobic conditions, K305M MAO A was normally reduced by substrate, whereas reoxidation by O2 was much slower but could be accomplished by quinone electron acceptors. When overexpressed in cardiomyoblasts by adenoviral vectors, the K305M variant showed enzymatic turnover similar to that of the wild-type but displayed decreased ROS levels and senescence markers. These results might translate into pharmacological treatments as MAO inhibitors may attenuate cardiomyocytes aging.

中文翻译:

将单胺氧化酶A合理地重新设计为脱氢酶以探测心脏衰老中的ROS。

心脏衰老是老年人群中典型的慢性衰弱病,细胞衰老通常与氧化应激有关。线粒体膜黄素酶单胺氧化酶A(MAO A)催化神经递质的氧化脱氨作用,并在老年心脏中增加其表达。我们在Lys305生产了重组人MAO A变体,它们在导致H 2 O 2产生的O 2反应性中起关键作用。K305Q变体的活性与野生型酶相同,而K305M和K305S的k cat值低200倍和100倍,K m相似。在厌氧条件下,K305M MAO A通常被底物还原,而被O重新氧化2慢得多,但可以通过醌电子受体来完成。当腺病毒载体在心肌母细胞中过表达时,K305M变体显示出与野生型相似的酶促转化,但显示出降低的ROS水平和衰老标记。这些结果可能会转化为药物治疗,因为MAO抑制剂可能会减弱心肌细胞的衰老。
更新日期:2020-07-17
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