Measurement of H2O2 within living Drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix.
Measurement of H2O2 within living Drosophila during aging using a ratiometric mass spectrometry probe targeted to the mitochondrial matrix.
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DOI：
10.1016/j.cmet.2011.02.003
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发表时间：
2011-03-02
期刊：
影响因子：
29
通讯作者：
Murphy MP
中科院分区：
文献类型：
--
作者：
Cochemé HM;Quin C;McQuaker SJ;Cabreiro F;Logan A;Prime TA;Abakumova I;Patel JV;Fearnley IM;James AM;Porteous CM;Smith RA;Saeed S;Carré JE;Singer M;Gems D;Hartley RC;Partridge L;Murphy MP
过氧化氢（H2 O2）是线粒体氧化损伤和氧化还原信号的核心，但由于难以在体内测量线粒体H2 O2，因此对其作用了解甚少。在这里，我们报告了一个比率质谱探针的方法来评估体内线粒体基质H2 O2水平。探针MitoB包含驱动其在线粒体内积累的三苯基膦（TPP）阳离子，其与芳基硼酸缀合，芳基硼酸与H2 O2反应形成苯酚MitoP。通过液相色谱-串联质谱法定量MitoP/MitoB比率能够测量线粒体H2 O2的加权平均值，其主要报告活蝇内的胸肌线粒体。有一个增加线粒体H2 O2与年龄的苍蝇，这是不协调改变干预措施，调制寿命。我们的研究结果提供了研究体内线粒体ROS的方法，并表明虽然整体线粒体H2 O2的增加与衰老相关，但可能不是原因。一种靶向线粒体的质谱探针测量体内线粒体H2 O2总体线粒体H2 O2随着年龄的增长而增加，但可能与寿命无关增加的体力活动导致线粒体H2 O2减少依赖于总体线粒体ROS的假设现在可以在体内进行评估
Hydrogen peroxide (H2O2) is central to mitochondrial oxidative damage and redox signaling, but its roles are poorly understood due to the difficulty of measuring mitochondrial H2O2 in vivo. Here we report a ratiometric mass spectrometry probe approach to assess mitochondrial matrix H2O2 levels in vivo. The probe, MitoB, comprises a triphenylphosphonium (TPP) cation driving its accumulation within mitochondria, conjugated to an arylboronic acid that reacts with H2O2 to form a phenol, MitoP. Quantifying the MitoP/MitoB ratio by liquid chromatography-tandem mass spectrometry enabled measurement of a weighted average of mitochondrial H2O2 that predominantly reports on thoracic muscle mitochondria within living flies. There was an increase in mitochondrial H2O2 with age in flies, which was not coordinately altered by interventions that modulated life span. Our findings provide approaches to investigate mitochondrial ROS in vivo and suggest that while an increase in overall mitochondrial H2O2 correlates with aging, it may not be causative. ► A mitochondria-targeted mass spectrometry probe measures mitochondrial H2O2 in vivo ► Overall mitochondrial H2O2 increases with age but can be independent of life span ► Increased physical activity leads to a decrease in mitochondrial H2O2 ► Hypotheses dependent on overall mitochondrial ROS can now be assessed in vivo
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DOI：
10.1042/bj20081386
发表时间：
2009-01-01
期刊：
The Biochemical journal
影响因子：
--
作者：
Murphy MP
通讯作者：
Murphy MP
影响因子：
3.7
作者：
Brown, Anthony E.;Baumbach, Janina;Ligoxygakis, Petros
通讯作者：
Ligoxygakis, Petros
影响因子：
7.4
作者：
Qian, SY;Kadiiska, MB;Mason, RP
通讯作者：
Mason, RP
影响因子：
16.6
作者：
Murphy MP;Partridge L
通讯作者：
Partridge L
影响因子：
14.8
作者：
Miller, Evan W.;Tulyanthan, Orapim;Chang, Christopher J.
通讯作者：
Chang, Christopher J.