黏蛋白核心蛋白的质谱分析
Mass spectrometric analysis of mucin core proteins.
作者信息
Kesimer Mehmet, Sheehan John K
机构信息
Department of Biochemistry and Biophysics Cystic Fibrosis/Pulmonary Research Center, University of North Carolina, 4021 Thurston Bowles Bldg. CB#7248, Chapel Hill, NC, USA.
出版信息
Methods Mol Biol. 2012;842:67-79. doi: 10.1007/978-1-61779-513-8_4.
Abstract
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.
摘要
由于粘蛋白具有高度糖基化的性质（高达90%）、高分子量（200 kDa - 200 MDa）和尺寸（回转半径10 - 300 nm），通过蛋白质组学应用对其进行鉴定和表征具有一定难度。它们的核心蛋白极大且被寡糖高度取代，这使得仅能触及到其蛋白质的高度受限部分。因此，传统的基于一维或二维聚丙烯酰胺凝胶的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，我们介绍了当前采用改良鸟枪法蛋白质组学方法来鉴定这些复杂糖蛋白的方案。
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