TY - JOUR
T1 - Effect of chronic morphine exposure on the synaptic plasma-membrane subproteome of rats
T2 - A quantitative protein profiling study based on isotope-coded affinity tags and liquid chromatography/mass spectrometry
AU - Prokai, Laszlo
AU - Zharikova, Alevtina D.
AU - Stevens, Stanley M.
PY - 2005/2
Y1 - 2005/2
N2 - 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.
AB - 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.
KW - Drug abuse
KW - Electrospray ionization
KW - Isotope-coded affinity tag
KW - Liquid chromatography/mass spectrometry
KW - Morphine
KW - Proteomics
KW - Quantitative protein profiling
KW - Subproteome
KW - Synaptic plasma membrane
KW - Tandem mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=14744280985&partnerID=8YFLogxK
U2 - 10.1002/jms.736
DO - 10.1002/jms.736
M3 - Article
C2 - 15706614
AN - SCOPUS:14744280985
SN - 1076-5174
VL - 40
SP - 169
EP - 175
JO - Journal of Mass Spectrometry
JF - Journal of Mass Spectrometry
IS - 2
ER -