Enrichment of low-abundance brain proteins by preparative electrophoresis.
Detection of low-copy-number gene products is essential for the development of novel drugs, however, it represents a major drawback of proteomics and simultaneously a scientific challenge. We studied the enrichment of rat brain cytosolic proteins by preparative electrophoresis using the PrepCell apparatus. The electrophoresis was performed in the presence of 0.1% lithium dodecyl sulfate. The proteins eluted from the gel were analyzed by two-dimensional gel electrophoresis and identified by matrix-assisted laser desorption ionization mass specrometry. Lithium dodecyl sulfate was easily exchanged against agents compatible with isoelectric focusing. Low-abundance proteins, which had not been found before, including neuronal-specific and calcium-binding proteins, were detected. In particular, low-molecular-mass proteins, such as hippocalcin, visinin-like proteins, and 14-3-3 proteins were strongly enriched by preparative electrophoresis.
Fountoulakis
Michael
M
F. Hoffmann-La Roche Ltd., Center for Medical Genomics, Basel, Switzerland. michael.fountoulakis@roche.com
Juranville
Jean François
JF
D000818:Animals; D001923:Brain Chemistry; D003600:Cytosol / Q000737:chemistry; D004586:Electrophoresis / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000302:isolation & purification*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009419:Nerve Tissue Proteins; D012967:Sodium Dodecyl Sulfate; C028913:dodecyl sulfate