An optimised method for the proteomic profiling of full thickness human skin.
An optimised method for the proteomic profiling of full thickness human skin.
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DOI：
10.1186/s12575-016-0045-y
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发表时间：
2016
影响因子：
6.4
通讯作者：
Mills K
中科院分区：
文献类型：
--
作者：
Bliss E;Heywood WE;Benatti M;Sebire NJ;Mills K
皮肤是人体最大的器官，是抵御外伤、微生物渗透和辐射的第一道屏障。皮肤病可能是多系统疾病或孤立病症的结果。由于其抗蛋白水解特性，与其他人体器官或体液相比，发表的人类皮肤蛋白质组数据集相对较少。由于皮肤的脂质含量高、不溶性和蛋白质广泛交联，因此使用传统蛋白质组技术分析皮肤是一种具有挑战性的组织。这会使使用质谱技术进行分析的蛋白质分离和消化变得复杂。我们优化了样品制备程序，以提高全层皮肤样品的溶解度和质谱兼容性。利用这项技术，我们能够使用在线二维液相色谱，然后进行超高清无标记质谱分析（UDMSE）来获得全层人类皮肤的蛋白质组图谱数据。我们能够从全层皮肤样本中识别出超过 2000 种蛋白质。采用离子淌度分离 (IMS) 技术的在线分级分离和优化采集方案，将蛋白质鉴定的范围显着扩大了十倍。本文的在线版本 (doi:10.1186/s12575-016-0045-y) 包含补充材料，可供授权用户使用。
The skin is the largest organ of the human body and is the first line barrier defence against trauma, microbial infiltration and radiation. Skin diseases can be a result of multi-systemic disease or an isolated condition. Due to its proteolysis resistant properties there are relatively few human skin proteomic datasets published compared with other human organs or body fluids. Skin is a challenging tissue to analyse using traditional proteomic techniques due to its high lipid content, insolubility and extensive cross-linking of proteins. This can complicate the isolation and digestion of proteins for analysis using mass spectrometry techniques. We have optimised a sample preparation procedure to improve solubilisation and mass spectral compatibility of full thickness skin samples. Using this technique, we were able to obtain data for the proteome profile of full thickness human skin using on-line two-dimensional liquid chromatography, followed by ultra-high definition label-free mass spectrometry analysis (UDMSE). We were able to identify in excess of 2000 proteins from a full thickness skin sample. The adoption of on-line fractionation and optimised acquisition protocols utilising ion mobility separation (IMS) technology has significantly increased the scope for protein identifications ten-fold. The online version of this article (doi:10.1186/s12575-016-0045-y) contains supplementary material, which is available to authorized users.