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Monitoring Membrane Lipidome Turnover by Metabolic 15N Labeling and Shotgun Ultra-High-Resolution Orbitrap Fourier Transform Mass Spectrometry
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-22 00:00:00 , DOI: 10.1021/acs.analchem.7b03437 Kai Schuhmann 1 , Kristina Srzentić 2 , Konstantin O. Nagornov 3 , Henrik Thomas 1 , Theresia Gutmann 4, 5 , Ünal Coskun 4, 5 , Yury O. Tsybin 3 , Andrej Shevchenko 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2017-11-22 00:00:00 , DOI: 10.1021/acs.analchem.7b03437 Kai Schuhmann 1 , Kristina Srzentić 2 , Konstantin O. Nagornov 3 , Henrik Thomas 1 , Theresia Gutmann 4, 5 , Ünal Coskun 4, 5 , Yury O. Tsybin 3 , Andrej Shevchenko 1
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Lipidomes undergo permanent extensive remodeling, but how the turnover rate differs between lipid classes and molecular species is poorly understood. We employed metabolic 15N labeling and shotgun ultra-high-resolution mass spectrometry (sUHR) to quantify the absolute (molar) abundance and determine the turnover rate of glycerophospholipids and sphingolipids by direct analysis of total lipid extracts. sUHR performed on a commercial Orbitrap Elite instrument at the mass resolution of 1.35 × 106 (m/z 200) baseline resolved peaks of 13C isotopes of unlabeled and monoisotopic peaks of 15N labeled lipids (Δm = 0.0063 Da). Therefore, the rate of metabolic 15N labeling of individual lipid species could be determined without compromising the scope, accuracy, and dynamic range of full-lipidome quantitative shotgun profiling. As a proof of concept, we employed sUHR to determine the lipidome composition and fluxes of 62 nitrogen-containing membrane lipids in human hepatoma HepG2 cells.
中文翻译：
通过代谢15 N标记和Shot弹枪超高分辨率Orbitrap傅里叶变换质谱监测膜脂质组转换
脂质组经历了永久性的广泛重塑，但是人们对脂质种类和分子种类之间的转换率如何不同的了解很少。我们采用了代谢15 N标记和shot弹枪超高分辨率质谱（sUHR）来定量绝对（摩尔）丰度，并通过直接分析总脂质提取物来确定甘油磷脂和鞘脂的周转率。苏尔在1.35×10质量分辨率以商业的Orbitrap精英仪器上进行6（米/ Ž 200）基线分辨的峰13个的未标记的和单一同位素峰Ç同位素15 Ñ标记的脂质（Δ米= 0.0063道尔顿）。因此，新陈代谢的速度可以确定单个脂质种类的15 N标记，而不会影响全脂质组定量shot弹枪分析的范围，准确性和动态范围。作为概念的证明，我们使用了sUHR来测定人肝癌HepG2细胞中的脂质组组成和62种含氮膜脂质的通量。
更新日期：2017-11-23
中文翻译：
通过代谢15 N标记和Shot弹枪超高分辨率Orbitrap傅里叶变换质谱监测膜脂质组转换
脂质组经历了永久性的广泛重塑，但是人们对脂质种类和分子种类之间的转换率如何不同的了解很少。我们采用了代谢15 N标记和shot弹枪超高分辨率质谱（sUHR）来定量绝对（摩尔）丰度，并通过直接分析总脂质提取物来确定甘油磷脂和鞘脂的周转率。苏尔在1.35×10质量分辨率以商业的Orbitrap精英仪器上进行6（米/ Ž 200）基线分辨的峰13个的未标记的和单一同位素峰Ç同位素15 Ñ标记的脂质（Δ米= 0.0063道尔顿）。因此，新陈代谢的速度可以确定单个脂质种类的15 N标记，而不会影响全脂质组定量shot弹枪分析的范围，准确性和动态范围。作为概念的证明，我们使用了sUHR来测定人肝癌HepG2细胞中的脂质组组成和62种含氮膜脂质的通量。
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