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Quantitative peptidomics in mice

Effect of cocaine treatment

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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 Cpe fat/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 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.

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Authors and Affiliations

  1. Department of Molecular Pharmacology, Albert Einstein College of Medicine, 10461, Bronx, NY

    Fa-Yun Che & Lloyd D. Fricker

  2. Department of Psychiatry and Behavioral Sciences, Albert Einstein College of Medicine, 10461, Bronx, NY

    Ilona Vathy

  3. Department of Neuroscience, Albert Einstein College of Medicine, 10461, Bronx, NY

    Ilona Vathy & Lloyd D. Fricker

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  1. Fa-Yun Che
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  2. Ilona Vathy
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Correspondence to Lloyd D. Fricker.

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Che, FY., Vathy, I. & Fricker, L.D. Quantitative peptidomics in mice. J Mol Neurosci 28, 265–275 (2006). https://doi.org/10.1385/JMN:28:3:265

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  • DOI: https://doi.org/10.1385/JMN:28:3:265

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