Direct profiling of proteins in biological tissue sections by MALDI mass spectrometry
Direct profiling of proteins in biological tissue sections by MALDI mass spectrometry
复制标题
DOI：
10.1021/ac990781q
复制
发表时间：
1999-12-01
影响因子：
7.4
通讯作者：
Caprioli, RM
中科院分区：
文献类型：
--
作者：
Chaurand, P;Stoeckli, M;Caprioli, RM
使用基质辅助激光解吸电离（MALDI）质谱法（MS）完成了小鼠几个器官组织切片中蛋白质的直接分析。将新鲜组织切片并吸干在导电聚乙烯膜上。将干燥的膜印迹用基质（通常为芥子酸）包被，并在质谱仪中直接分析。通常，观察到在2000-30 000 Ha范围内远远超过100个肽/蛋白质信号，其中30-50个具有相对高的信号强度。同一组织的不同区域的分析给出了显著相似的质谱，具有大于90%的同源性，然而，分段组织的不同部分，例如近端、中间和远端结肠，给出了一些独特的蛋白质信号。用蛋白酶处理组织印迹并随后使用源后衰变方法进行肽测序进行MALDI MS分析后，鉴定了一些蛋白质。从这些组织印迹中测量的独特蛋白质谱也显示出小鼠品系之间的差异，遗传相似的品系具有非常相似的模式。
The direct profiling of proteins present in tissue sections for several organs of the mouse has been accomplished using matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS). Fresh tissue was sectioned and blotted on a conductive polyethylene membrane. The dried membrane blot was coated with matrix, typically sinapinic acid, and directly analyzed in the mass spectrometer, Generally, well over 100 peptide/protein signals in the 2000-30 000 Ha range were observed, with 30-50 having relatively high signal intensities. Analysis of different areas of the same tissue gave remarkably similar mass spectra with greater than 90% homology, However, different parts of a segmented tissue, such as the proximal, intermediate, and distal colon, gave some unique protein signals. After treatment of the tissue blot with protease and subsequent MALDI MS analysis using postsource decay methods for peptide sequencing, some of the proteins were identified. The unique protein profiles measured from these tissue blots also showed differences from strain to strain of the mouse, with genetically similar strains having very similar patterns.