Department of Chemistry, University of Wisconsin, Madison, Wisconsin 53706, United States.
Anal Chem. 2010 Dec 15;82(24):10068-74. doi: 10.1021/ac1020358. Epub 2010 Nov 9.
Using a modified electron transfer dissociation (ETD)-enabled quadrupole linear ion trap (QLT) mass spectrometer, we demonstrate the utility of IR activation concomitant with ETD ion-ion reactions (activated-ion ETD, AI-ETD). Analyzing 12 strong cation exchanged (SCX) fractions of a LysC digest of human cell protein extract using ETD, collision-activated dissociation (CAD), and AI-ETD, we find that AI-ETD generates 13 405 peptide spectral matches (PSMs) at a 1% false-discovery rate (1% FDR), surpassing both ETD (7 968) and CAD (10 904). We also analyze 12 SCX fractions of a tryptic digest of human cell protein extract and find that ETD produces 6 234 PSMs, AI-ETD 9 130 PSMs, and CAD 15 209 PSMs. Compared to ETD with supplemental collisional activation (ETcaD), AI-ETD generates ∼80% more PSMs for the whole cell lysate digested with trypsin and ∼50% more PSMs for the whole cell lysate digested with LysC.
利用改进的电子转移解离（ETD）功能的四极线性离子阱（QLT）质谱仪，我们展示了 IR 激活伴随 ETD 离子-离子反应（活化离子 ETD，AI-ETD）的实用性。通过 ETD、碰撞激活解离（CAD）和 AI-ETD 分析人细胞蛋白提取物 LysC 消化物的 12 个强阳离子交换（SCX）级分，我们发现 AI-ETD 在 1%假发现率（1% FDR）下生成 13405 个肽谱匹配（PSM），超过 ETD（7968）和 CAD（10904）。我们还分析了人细胞蛋白提取物胰蛋白酶消化物的 12 个 SCX 级分，发现 ETD 产生 6234 个 PSM，AI-ETD 产生 9130 个 PSM，CAD 产生 15209 个 PSM。与带有补充碰撞激活（ETcaD）的 ETD 相比，AI-ETD 对胰蛋白酶消化的整个细胞裂解物产生的 PSM 增加了约 80%，对 LysC 消化的整个细胞裂解物产生的 PSM 增加了约 50%。
