Enrichment of low-abundance brain proteins by preparative electrophoresis

Anal Biochem. 2003 Feb 15;313(2):267-82. doi: 10.1016/s0003-2697(02)00617-6.

Abstract

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.

MeSH terms

  • Animals
  • Brain Chemistry*
  • Cytosol / chemistry
  • Electrophoresis / methods
  • Electrophoresis, Gel, Two-Dimensional
  • Molecular Weight
  • Nerve Tissue Proteins / isolation & purification*
  • Proteomics
  • Rats
  • Rats, Sprague-Dawley
  • Sodium Dodecyl Sulfate / chemistry
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Nerve Tissue Proteins
  • Sodium Dodecyl Sulfate
  • dodecyl sulfate