In-depth proteomic analysis of mouse cochlear sensory epithelium by mass spectrometry.
In-depth proteomic analysis of mouse cochlear sensory epithelium by mass spectrometry.
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DOI：
10.1021/pr4001338
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发表时间：
2013-08-02
影响因子：
4.4
通讯作者：
Sokolowski BH
中科院分区：
文献类型：
--
作者：
Darville LN;Sokolowski BH
感觉器官如耳蜗的蛋白质组学分析是具有挑战性的，因为它的体积小，膜蛋白分离困难。质谱法结合分离方法可以提供更全面的蛋白质组，因为它能够丰富蛋白质样品，检测疏水性蛋白质，并通过减少蛋白质组动态范围来识别低丰度蛋白质。GELFrEE以及不同的分离和消化技术结合FASP和nanoLC-MS/MS对30日龄小鼠耳蜗感觉上皮进行了深入的蛋白质组分析。用LysC/trypsin酶切，然后用SCX分离和多次nanoLC-MS/MS分析，鉴定出3773个蛋白，FDR为1%。其中，694个蛋白id位于质膜中。将GELFrEE/LysC/trypsin的结果与GELFrEE/trypsin/trypsin的结果结合得到的蛋白id产生2779个蛋白，其中606个蛋白使用GELFrEE/LysC/trypsin方法鉴定。结合不同技术的结果，总共得到了4620个id，包括一些以前未报道的蛋白质。氧化石墨烯分析显示，结合蛋白和催化蛋白以及与代谢相关的蛋白高表达。结果表明，需要多种技术的应用，以提供一个详尽的蛋白质组的耳蜗感觉上皮，包括许多膜蛋白。质谱蛋白质组学数据已存入ProteomeXchange Consortium，数据集标识符为PXD000231。
Proteomic analysis of sensory organs such as the cochlea is challenging due to its small size and difficulties with membrane protein isolation. Mass spectrometry in conjunction with separation methods can provide a more comprehensive proteome, because of the ability to enrich protein samples, detect hydrophobic proteins, and identify low abundant proteins by reducing the proteome dynamic range. GELFrEE as well as different separation and digestion techniques were combined with FASP and nanoLC-MS/MS to obtain an in-depth proteome analysis of cochlear sensory epithelium from 30-day-old mice. Digestion with LysC/trypsin followed by SCX fractionation and multiple nanoLC-MS/MS analyses identified 3773 proteins with a 1% FDR. Of these, 694 protein IDs were in the plasmalemma. Protein IDs obtained by combining outcomes from GELFrEE/LysC/trypsin with GELFrEE/trypsin/trypsin generated 2779 proteins, of which 606 additional proteins were identified using the GELFrEE/LysC/trypsin approach. Combining results from the different techniques resulted in a total of 4620 IDs, including a number of previously unreported proteins. GO analyses showed high expression of binding and catalytic proteins as well as proteins associated with metabolism. The results show that the application of multiple techniques is needed to provide an exhaustive proteome of the cochlear sensory epithelium that includes many membrane proteins. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium with the data set identifier PXD000231.
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影响因子：
3.7
作者：
Elkan-Miller T;Ulitsky I;Hertzano R;Rudnicki A;Dror AA;Lenz DR;Elkon R;Irmler M;Beckers J;Shamir R;Avraham KB
通讯作者：
Avraham KB
影响因子：
5.3
作者：
Gagnon, Leona H.;Longo-Guess, Chantal M.;Johnson, Kenneth R.
通讯作者：
Johnson, Kenneth R.
DOI：
10.1073/pnas.0600923103
发表时间：
2006-07-18
影响因子：
11.1
作者：
Mburu, Philomena;Kikkawa, Yoshiaki;Brown, Steve D. M.
通讯作者：
Brown, Steve D. M.
影响因子：
3.3
作者：
Jamesdaniel S;Hu B;Kermany MH;Jiang H;Ding D;Coling D;Salvi R
通讯作者：
Salvi R
影响因子：
5.5
作者：
Marcotti, W;Johnson, SL;Kros, CJ
通讯作者：
Kros, CJ