Targeting mitochondria with small molecules: the preparation of MitoB and MitoP as exomarkers of mitochondrial hydrogen peroxide.
Targeting mitochondria with small molecules: the preparation of MitoB and MitoP as exomarkers of mitochondrial hydrogen peroxide.
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DOI：
10.1007/978-1-4939-2288-8_3
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发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Hartley, Richard C
中科院分区：
文献类型：
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作者：
Cairns, Andrew G;McQuaker, Stephen J;Hartley, Richard C
使用亲脂性烷基三苯基鏻 (TPP) 基团可以将小分子以物理化学方式靶向线粒体。一旦进入线粒体，TPP-缀合物就可以直接检测或影响线粒体基质内的过程。或者，缀合物可以作为前药，通过使用内部或外部指令从 TPP 基团释放来激活。可以设计可用于任何细胞系、组织或整个生物体的小分子，允许时间控制，并以可逆的剂量依赖性方式应用。一个例子是 MitoB 对整个生物体线粒体中过氧化氢的检测和定量。线粒体基质内产生的过氧化氢参与信号传导，并涉及与衰老和多种与年龄相关的疾病（包括心血管疾病、神经退行性变和癌症）相关的氧化损伤。 MitoB 在线粒体中积累，并通过线粒体基质中的过氧化氢转化为外切标记物 MitoP。过氧化氢浓度由孵育一段时间后MitoP与MitoB的比率确定，并且该比率是使用d15-MitoP和d15-MitoB作为标准通过质谱法测定的。在这里，我们描述了 MitoB 和 MitoP 的合成以及这种定量方法所需的氘代标准品。
Small molecules can be physicochemically targeted to mitochondria using the lipophilic alkyltriphenylphosphonium (TPP) group. Once in the mitochondria the TPP-conjugate can detect or influence processes within the mitochondrial matrix directly. Alternatively, the conjugate can behave as a prodrug, which is activated by release from the TPP group either using an internal or external instruction. Small molecules can be designed that can be used in any cell line, tissue or whole organism, allow temporal control, and be applied in a reversible dose-dependent fashion. An example is the detection and quantification of hydrogen peroxide in mitochondria of whole living organisms by MitoB. Hydrogen peroxide produced within the mitochondrial matrix is involved in signalling and implicated in the oxidative damage associated with aging and a wide range of age-associated conditions including cardiovascular disease, neurodegeneration, and cancer. MitoB accumulates in mitochondria and is converted into the exomarker, MitoP, by hydrogen peroxide in the mitochondrial matrix. The hydrogen peroxide concentration is determined from the ratio of MitoP to MitoB after a period of incubation, and this ratio is determined by mass spectrometry using d15-MitoP and d15-MitoB as standard. Here we describe the synthesis of MitoB and MitoP and the deuterated standards necessary for this method of quantification.