Effect of chronic morphine exposure on the synaptic plasma-membrane subproteome of rats: A quantitative protein profiling study based on isotope-coded affinity tags and liquid chromatography/mass spectrometry

Laszlo Prokai, Alevtina D. Zharikova, Stanley M. Stevens

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

The effect of chronic morphine exposure on the synaptic plasma-membrane subproteome in rats was studied by the isotope-coded affinity tag (ICAT) method coupled with capillary reversed-phase liquid chromatography/electrospray ionization mass spectrometry and tandem mass spectrometry. ICAT-labeled tryptic peptides of synaptic membrane proteins were successfully identified using tandem mass spectrometry in conjunction with protein database searching. Several important synaptic plasma-membrane proteins displayed significant regulation changes as a result of chronic morphine exposure in vivo. In particular, an integral membrane protein Na+/IC+ ATPase (α-subunit) involved in regulation of the cell membrane potential by controlling sodium and potassium ion permeability was downregulated by 39 ± 2%. This result was in excellent agreement with the reduction in electrogenic Na+, K+ pumping due to about 40% downregulation of Na+TK + ATPase α3-isoform in myenteric S-neurons of morphine-exposed guinea-pigs measured by others via immunohistochemistry. The decrease in the abundance of non-erythroid αII-spectrin in the synaptic plasma-membrane fraction was also observed, which was hypothetically associated with the breakdown of the protein due to the upregulation of the proteolytic enzyme caspase-3 upon chronic morphine exposure.

Original languageEnglish
Pages (from-to)169-175
Number of pages7
JournalJournal of Mass Spectrometry
Volume40
Issue number2
DOIs
StatePublished - Feb 2005

Keywords

  • Drug abuse
  • Electrospray ionization
  • Isotope-coded affinity tag
  • Liquid chromatography/mass spectrometry
  • Morphine
  • Proteomics
  • Quantitative protein profiling
  • Subproteome
  • Synaptic plasma membrane
  • Tandem mass spectrometry

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