Chaurand P, Stoeckli M, Caprioli R M
Mass Spectrometry Research Center, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6400, USA.
Anal Chem. 1999 Dec 1;71(23):5263-70. doi: 10.1021/ac990781q.
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 Da 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.
利用基质辅助激光解吸电离(MALDI)质谱(MS)技术,已实现对小鼠多个器官组织切片中存在的蛋白质进行直接分析。将新鲜组织切片并印迹在导电聚乙烯膜上。干燥后的膜印迹用基质(通常为芥子酸)包被,然后直接在质谱仪中进行分析。一般来说,在2000 - 30000 Da范围内可观察到远超100个肽/蛋白质信号,其中30 - 50个具有相对较高的信号强度。对同一组织的不同区域进行分析,得到的质谱图非常相似,同源性大于90%。然而,分段组织的不同部分,如近端、中间和远端结肠,会产生一些独特的蛋白质信号。在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的MALDI MS分析后,鉴定出了一些蛋白质。从这些组织印迹中测得的独特蛋白质谱在不同品系的小鼠之间也存在差异,基因相似的品系具有非常相似的模式。