Proteomic analysis of the fetal brain.
We applied proteomic technologies to analyze the human fetal brain. Such an analysis could provide us with important information on the development of the early neuronal life in healthy and diseased states. The proteins from the cerebellum of control subjects were analyzed by two-dimensional electrophoresis and identified by matrix-assisted laser desorption/ionization-mass spectrometry on the basis of peptide mass fingerprinting, following in-gel digestion with trypsin. Approximately 3,000 spots, excised from three two-dimensional gels, were analyzed which resulted in the identification of about 1,700 proteins that were the products of 437 different genes. About half of them are enzyme subunits and are mainly localized in the cytosol and in mitochondria. The most frequently identified proteins in the various gels were heat shock proteins, house-keeping enzymes, such as ATP synthase chains, protein disulfide isomerase, and structural proteins, such as tubulin chains. Seven gene products were identified for the first time in the fetal brain. The other proteins had also been detected in other human samples which were analyzed in our laboratory. Most proteins were represented by multiple spots. In average, about 3-5 spots were detected per gene product. The fetal brain database includes proteins with important functions and also with unknown functions and represents today one of the largest two-dimensional databases for higher eukaryotic proteomes. It may be a useful tool in the investigation of protein changes in neurodegenerative diseases early in life.
Fountoulakis
Michael
M
Center for Medical Genomics, F. Hoffmann-La Roche, Basel, Switzerland. michael.fountoulakis@roche.com
Juranville
Jean-François
JF
Dierssen
Mara
M
Lubec
Gert
G
D001921:Brain / Q000196:embryology* / Q000378:metabolism*; D001923:Brain Chemistry; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional; D005333:Fetus / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D020543:Proteome / Q000032:analysis / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D020543:Proteome