Changes in the levels of low-abundance brain proteins induced by kainic acid.
Low-abundance gene products are of interest in proteomic studies, because they are probably involved in disease-related changes and their altered levels or modifications may carry significant biological information. Detection of low-abundance proteins of a proteome is one of the major limitations of proteomics and a scientific challenge. We investigated the changes in the levels of low-abundance rat brain cytosolic proteins after administration of kainic acid, a potent neurotoxin and excitatory amino acid. The cytosolic proteins from controls and animals treated with kainic acid were fractionated on an ion-exchange column. The fractions collected were analyzed by 2D electrophoresis, and the proteins with altered levels were identified by matrix-assisted laser desorption ionization or ion-spray MS. We found a manifold decrease in annexin VII, heat-shock cofactor HOP/p60 and SP-22 and a manifold increase in heparin-binding protein p30. The results suggest, respectively, the involvement of an apoptotic pathway, recruitment of the heat-shock protein machinery, generation of an antioxidant response, and, probably, induction of repair mechanisms. Three of the four proteins with altered levels had not been previously detected in the cytosolic fraction, and detection of the altered levels was possible only after the protein-enriching step.
Krapfenbauer
K
K
F. Hoffmann-La Roche Ltd, Pharmaceutical Research, Genomics Technologies, University of Vienna, Austria.
Berger
M
M
Friedlein
A
A
Lubec
G
G
Fountoulakis
M
M
D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D015180:Electrophoresis, Gel, Two-Dimensional; D007608:Kainic Acid / Q000494:pharmacology*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D009419:Nerve Tissue Proteins; D007608:Kainic Acid