Abstract
We recently developed a quantitative peptidomics method using stable isotopic labels and mass spectrometry to both quantify and identify a large number of peptides. To test this approach and screen for peptides regulated by cocaine administration, 32 Cpefat/fat mice and 16 wild-type mice were treated twice daily for 5 d either with saline or 10 mg/kg cocaine. Peptides were extracted from striatum, hypothalamus, hippocampus, and prefrontal cortex, and extracts from groups of eight mice were labeled with the N-hydroxysuccinimide ester of trimethylammonium butyrate containing either nine deuterium or nine hydrogen atoms. Pools of heavy- and light-labeled peptides were combined, purified on an anhydrotrypsin affinity column, and analyzed on a reverse-phase column coupled to an electrospray ionization quadrapole time-of-flight mass spectrometer. Changes in peptide levels upon cocaine treatment were determined from the relative peak intensities of the cocaine versus saline peaks, and peptides were identified from collision-induced dissociation spectra. Ten peptides were found to increase or decrease in each of two separate analyses from distinct groups of mice. Peptides found to increase corresponded to fragments of proenkephalin, prothyrotropin-releasing hormone, provasopressin, proSAAS, secretogranin II, chromogranin B, and peptidyl-glycine-α-amidating mono-oxygenase in the hypothalamus. The same peptidyl-glycine-α-amidating mono-oxygenase peptide decreased in the prefrontal cortex, along with striatal neurokinin B and two unidentified peptides. Thirty other peptides were not substantially affected by cocaine treatment in both replicates. Taken together, the quantitative peptidomics approach provides an efficient method to screen for changes in a large number of peptides.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 265-275 |
| Number of pages | 11 |
| Journal | Journal of Molecular Neuroscience |
| Volume | 28 |
| Issue number | 3 |
| DOIs | |
| State | Published - 2006 |
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Keywords
- Carboxypeptidase
- Peptide processing
- Preprotachykinin B
- Proenkephalin
- proSAAS
- Secretogranin II
ASJC Scopus subject areas
- Neuroscience(all)
- Biochemistry
- Genetics
Cite this
Quantitative peptidomics in mice : Effect of cocaine treatment. / Che, Fa Yun; Vathy, Ilona; Fricker, Lloyd D.
In: Journal of Molecular Neuroscience, Vol. 28, No. 3, 2006, p. 265-275.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Quantitative peptidomics in mice
T2 - Effect of cocaine treatment
AU - Che, Fa Yun
AU - Vathy, Ilona
AU - Fricker, Lloyd D.
PY - 2006
Y1 - 2006
N2 - We recently developed a quantitative peptidomics method using stable isotopic labels and mass spectrometry to both quantify and identify a large number of peptides. To test this approach and screen for peptides regulated by cocaine administration, 32 Cpefat/fat mice and 16 wild-type mice were treated twice daily for 5 d either with saline or 10 mg/kg cocaine. Peptides were extracted from striatum, hypothalamus, hippocampus, and prefrontal cortex, and extracts from groups of eight mice were labeled with the N-hydroxysuccinimide ester of trimethylammonium butyrate containing either nine deuterium or nine hydrogen atoms. Pools of heavy- and light-labeled peptides were combined, purified on an anhydrotrypsin affinity column, and analyzed on a reverse-phase column coupled to an electrospray ionization quadrapole time-of-flight mass spectrometer. Changes in peptide levels upon cocaine treatment were determined from the relative peak intensities of the cocaine versus saline peaks, and peptides were identified from collision-induced dissociation spectra. Ten peptides were found to increase or decrease in each of two separate analyses from distinct groups of mice. Peptides found to increase corresponded to fragments of proenkephalin, prothyrotropin-releasing hormone, provasopressin, proSAAS, secretogranin II, chromogranin B, and peptidyl-glycine-α-amidating mono-oxygenase in the hypothalamus. The same peptidyl-glycine-α-amidating mono-oxygenase peptide decreased in the prefrontal cortex, along with striatal neurokinin B and two unidentified peptides. Thirty other peptides were not substantially affected by cocaine treatment in both replicates. Taken together, the quantitative peptidomics approach provides an efficient method to screen for changes in a large number of peptides.
AB - We recently developed a quantitative peptidomics method using stable isotopic labels and mass spectrometry to both quantify and identify a large number of peptides. To test this approach and screen for peptides regulated by cocaine administration, 32 Cpefat/fat mice and 16 wild-type mice were treated twice daily for 5 d either with saline or 10 mg/kg cocaine. Peptides were extracted from striatum, hypothalamus, hippocampus, and prefrontal cortex, and extracts from groups of eight mice were labeled with the N-hydroxysuccinimide ester of trimethylammonium butyrate containing either nine deuterium or nine hydrogen atoms. Pools of heavy- and light-labeled peptides were combined, purified on an anhydrotrypsin affinity column, and analyzed on a reverse-phase column coupled to an electrospray ionization quadrapole time-of-flight mass spectrometer. Changes in peptide levels upon cocaine treatment were determined from the relative peak intensities of the cocaine versus saline peaks, and peptides were identified from collision-induced dissociation spectra. Ten peptides were found to increase or decrease in each of two separate analyses from distinct groups of mice. Peptides found to increase corresponded to fragments of proenkephalin, prothyrotropin-releasing hormone, provasopressin, proSAAS, secretogranin II, chromogranin B, and peptidyl-glycine-α-amidating mono-oxygenase in the hypothalamus. The same peptidyl-glycine-α-amidating mono-oxygenase peptide decreased in the prefrontal cortex, along with striatal neurokinin B and two unidentified peptides. Thirty other peptides were not substantially affected by cocaine treatment in both replicates. Taken together, the quantitative peptidomics approach provides an efficient method to screen for changes in a large number of peptides.
KW - Carboxypeptidase
KW - Peptide processing
KW - Preprotachykinin B
KW - Proenkephalin
KW - proSAAS
KW - Secretogranin II
UR - http://www.scopus.com/inward/record.url?scp=33646880017&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646880017&partnerID=8YFLogxK
U2 - 10.1385/JMN:28:3:265
DO - 10.1385/JMN:28:3:265
M3 - Article
C2 - 16691014
AN - SCOPUS:33646880017
VL - 28
SP - 265
EP - 275
JO - Journal of Molecular Neuroscience
JF - Journal of Molecular Neuroscience
SN - 0895-8696
IS - 3
ER -