Mass Spectrometric Analysis of Mucin Core Proteins
Mass Spectrometric Analysis of Mucin Core Proteins
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DOI：
10.1007/978-1-61779-513-8_4
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Sheehan, John K.
中科院分区：
文献类型：
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作者：
Kesimer, Mehmet;Sheehan, John K.
由于粘蛋白具有高度糖基化性质（高达 90%）、高分子量（200 kDa-200 MDa）和大小（Rg 10-300 nm），因此很难通过蛋白质组学应用对其进行鉴定和表征。它们的核心蛋白非常大，并且被寡糖高度取代，这使得只能访问其蛋白质的高度受限的部分。因此，传统的一维或二维聚丙烯酰胺凝胶蛋白质组学方法对于粘蛋白分子的鉴定和表征并不有效。在本章中，我们介绍了我们目前的方案，采用改良的鸟枪蛋白质组学方法来识别这些复杂的糖蛋白。
Mucins are difficult to handle for their identification and characterization via proteomic applications due to their heavily glycosylated nature (up to 90%), high molecular weight (200 kDa-200 MDa), and size (Rg 10-300 nm). Their core proteins are extremely large and highly substituted with oligosaccharides, which only allow access to a highly restricted portion of their protein. For this reason, conventional 1D or 2D polyacrylamide gel-based proteomic approaches are not effective for identification and characterization of mucin molecules. In this chapter, we present our current protocol employing a modified shotgun proteomic approach to identify these complex glycoproteins.