Carbene footprinting accurately maps binding sites in protein-ligand and protein-protein interactions.
Carbene footprinting accurately maps binding sites in protein-ligand and protein-protein interactions.
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DOI：
10.1038/ncomms13288
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发表时间：
2016-11-16
影响因子：
16.6
通讯作者：
Oldham, Neil J.
中科院分区：
文献类型：
--
作者：
Manzi, Lucio;Barrow, Andrew S.;Scott, Daniel;Layfield, Robert;Wright, Timothy G.;Moses, John E.;Oldham, Neil J.
蛋白质与其结合伙伴之间的特异性相互作用是生命过程的基础。检测蛋白质复合物并绘制其结合位点的能力对于在分子水平上理解基础生物学至关重要。采用灵敏分析技术（如质谱法）的方法有可能在很短的时间内用很少的材料提供有价值的见解。在这里，我们提出了一种差异蛋白质足迹技术，采用高效的光活化探针与质谱。使用这种方法，碳水化合物底物的位置被准确地映射到溶菌酶的结合裂缝，并且在一个更复杂的例子中，100 kDa的多结构域去泛素化酶USP 5和双泛素底物之间的相互作用被定位到不同的功能结构域。这种探针的大大改进的性质使得卡宾足迹法成为一种可行的方法，用于利用良性的近紫外光活化快速准确地鉴定蛋白质结合位点。 绘制蛋白质-配体相互作用是推进细胞过程理解和开发药物发现策略的基础。在这里，作者提出了一种光活化探针，可以使用质谱法高效标记和识别蛋白质结合位点。
Specific interactions between proteins and their binding partners are fundamental to life processes. The ability to detect protein complexes, and map their sites of binding, is crucial to understanding basic biology at the molecular level. Methods that employ sensitive analytical techniques such as mass spectrometry have the potential to provide valuable insights with very little material and on short time scales. Here we present a differential protein footprinting technique employing an efficient photo-activated probe for use with mass spectrometry. Using this methodology the location of a carbohydrate substrate was accurately mapped to the binding cleft of lysozyme, and in a more complex example, the interactions between a 100 kDa, multi-domain deubiquitinating enzyme, USP5 and a diubiquitin substrate were located to different functional domains. The much improved properties of this probe make carbene footprinting a viable method for rapid and accurate identification of protein binding sites utilizing benign, near-UV photoactivation. Mapping protein-ligand interactions is fundamental to advance the understanding of cellular processes and to develop drug discovery strategies. Here, the authors present a photo-activated probe that allows highly efficient labelling and identification of protein binding sites using mass spectrometry.
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DOI：
10.1016/j.ymeth.2015.02.018
发表时间：
2015-11-01
期刊：
Methods (San Diego, Calif.)
影响因子：
--
作者：
Calabrese AN;Ault JR;Radford SE;Ashcroft AE
通讯作者：
Ashcroft AE
DOI：
10.1039/a707586c
发表时间：
1998-05-01
期刊：
JOURNAL OF THE CHEMICAL SOCIETY-PERKIN TRANSACTIONS 2
影响因子：
--
作者：
Admasu, A;Gudmundsdottir, AD;Crocker, PJ
通讯作者：
Crocker, PJ
影响因子：
7.4
作者：
Gomez, Gabriela Elena;Monti, Jose Luis E.;Delfino, Jose Maria
通讯作者：
Delfino, Jose Maria
影响因子：
7.4
作者：
Jumper, Chanelle C.;Bomgarden, Ryan;Schriemer, David C.
通讯作者：
Schriemer, David C.
影响因子：
7.4
作者：
Aye, TT;Low, TY;Sze, SK
通讯作者：
Sze, SK