Detection and identification of 4-hydroxy-2-nonenal Schiff-base adducts along with products of Michael addition using data-dependent neutral loss-driven MS3 acquisition: method evaluation through an in vitro study on cytochrome c oxidase modifications.

Detection and identification of 4-hydroxy-2-nonenal Schiff-base adducts along with products of Michael addition using data-dependent neutral loss-driven MS3 acquisition: method evaluation through an in vitro study on cytochrome c oxidase modifications.
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DOI:
10.1002/pmic.200900116
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发表时间:
2009-11
期刊:
影响因子:
3.4
通讯作者:
Prokai, Laszlo
Prokai, Laszlo
中科院分区:
生物学3区
文献类型:
--
作者:
Rauniyar, Navin;Prokai, Laszlo

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我们报告了一个数据依赖的中性损失驱动的MS3获取,除了丰富的Michael加合物外,还增强了蛋白质和4-羟基-2-壬烯醛(脂质过氧化反应的最终产物)的希夫碱加合物的检测和定位。以线粒体蛋白复合物细胞色素c氧化酶体外修饰为模型,对该方法进行了评价。该技术允许对修饰位点进行可靠的验证,并在蛋白质复合物的Vb亚基中鉴定出希夫碱加合物。
We report a data-dependent neutral-loss driven MS3 acquisition to enhance, in addition to abundant Michael adducts, the detection and localization of Schiff-base adducts of proteins and 4-hydroxy-2-nonenal, a reactive end-products of lipid peroxidation. In vitro modification of cytochrome c oxidase, a mitochondrial protein complex, was used as a model to evaluate the method. The technique allowed for a confident validation of modification sites and also identified a Schiff-base adduct in subunit Vb of the protein complex.
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