Profiling proteins from azoxymethane-induced colon tumors at the molecular level by matrix-assisted laser desorption/ionization mass spectrometry.
New developments in mass spectrometry allow for the profiling of the major proteomic content of fresh tissue sections. Briefly, fresh tissue sections are sampled and blotted onto a polyethylene membrane for protein transfer and then subsequently analyzed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS). Using this technology, we have compared the protein expression of normal and cancerous mouse colon tissue obtained from the same animal. By difference, several protein signals specific to cancerous tissue were observed. A protein extract obtained from the tumors was fractionated by high-performance liquid chromatography and the individual fractions analyzed by MALDI-MS. The fractions containing the targeted proteins were subjected to trypsin digestion. The resulting tryptic peptides were sequenced by tandem mass spectrometry, and based on the recovered partial amino acid sequences, three of the tumor specific protein markers were identified as calgranulin A (S100A8), calgranulin B (S100A9) and calgizzarin (S100A11).
Chaurand
P
P
Mass Spectrometry Research Center, Department of Biochemistry, Vanderbilt University, 824A Robinson Research Building, Nashville, TN 37232-6400, USA.
DaGue
B B
BB
Pearsall
R S
RS
Threadgill
D W
DW
Caprioli
R M
RM
D000818:Animals; D001391:Azo Compounds / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D003110:Colonic Neoplasms / Q000139:chemically induced* / Q000378:metabolism*; D004195:Disease Models, Animal; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D051379:Mice; D009363:Neoplasm Proteins / Q000032:analysis; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001391:Azo Compounds; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; C001825:azomethane