Direct Profiling of Proteins in Biological Tissue Sections by MALDI Mass Spectrometry

• Published in: Analytical chemistry (Washington) 1999-12, Vol.71 (23), p.5263-5270
• Main Authors: Chaurand, Pierre, Stoeckli, Markus, Caprioli, Richard M
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Colon - chemistry; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Mice; Proteins; Proteins - chemistry; Rats; Reproducibility of Results; Rodents; Scientific imaging; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Tissues
• ISSN: 0003-2700; 1520-6882
• 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.