Fournier Frédéric, Gardner Elizabeth M, Kedra Darek A, Donaldson Paul M, Guo Rui, Butcher Sarah A, Gould Ian R, Willison Keith R, Klug David R
Department of Chemistry, Imperial College London, Exhibition Road, London SW7 2AZ, United Kingdom.
Proc Natl Acad Sci U S A. 2008 Oct 7;105(40):15352-7. doi: 10.1073/pnas.0805127105. Epub 2008 Oct 1.
Electron-vibration-vibration two-dimensional coherent spectroscopy, a variant of 2DIR, is shown to be a useful tool to differentiate a set of 10 proteins based on their amino acid content. Two-dimensional vibrational signatures of amino acid side chains are identified and the corresponding signal strengths used to quantify their levels by using a methyl vibrational feature as an internal reference. With the current apparatus, effective differentiation can be achieved in four to five minutes per protein, and our results suggest that this can be reduced to <1 min per protein by using the same technology. Finally, we show that absolute quantification of protein levels is relatively straightforward to achieve and discuss the potential of an all-optical high-throughput proteomic platform based on two-dimensional infrared spectroscopic measurements.
电子-振动-振动二维相干光谱（2DIR的一种变体）被证明是一种基于氨基酸含量区分一组10种蛋白质的有用工具。识别出氨基酸侧链的二维振动特征，并以甲基振动特征作为内部参考，利用相应的信号强度来量化它们的水平。使用当前仪器，每种蛋白质可在4至5分钟内实现有效区分，我们的结果表明，通过使用相同技术，这一时间可缩短至每种蛋白质<1分钟。最后，我们表明蛋白质水平的绝对定量相对容易实现，并讨论了基于二维红外光谱测量的全光学高通量蛋白质组学平台的潜力。
