Probing the production of amidated peptides following genetic and dietary copper manipulations

Ping Yin, Danielle Bousquet-Moore, Suresh P. Annangudi, Bruce R. Southey, Richard E. Mains, Betty A. Eipper, Jonathan V. Sweedler

Research output: Contribution to journalArticle

Abstract

Amidated neuropeptides play essential roles throughout the nervous and endocrine systems. Mice lacking peptidylglycine α-amidating monooxygenase (PAM), the only enzyme capable of producing amidated peptides, are not viable. In the amidation reaction, the reactant (glycine-extended peptide) is converted into a reaction intermediate (hydroxyglycine-extended peptide) by the copper-dependent peptidylglycine-α-hydroxylating monooxygenase (PHM) domain of PAM. The hydroxyglycine-extended peptide is then converted into amidated product by the peptidyl-α-hydroxyglycine α-amidating lyase (PAL) domain of PAM. PHM and PAL are stitched together in vertebrates, but separated in some invertebrates such as Drosophila and Hydra. In addition to its luminal catalytic domains, PAM includes a cytosolic domain that can enter the nucleus following release from the membrane by γ-secretase. In this work, several glycine- and hydroxyglycine-extended peptides as well as amidated peptides were qualitatively and quantitatively assessed from pituitaries of wild-type mice and mice with a single copy of the Pam gene (PAM +/-) via liquid chromatography-mass spectrometry-based methods. We provide the first evidence for the presence of a peptidyl-α-hydroxyglycine in vivo, indicating that the reaction intermediate becomes free and is not handed directly from PHM to PAL in vertebrates. Wild-type mice fed a copper deficient diet and PAM +/- mice exhibit similar behavioral deficits. While glycine-extended reaction intermediates accumulated in the PAM +/- mice and reflected dietary copper availability, amidated products were far more prevalent under the conditions examined, suggesting that the behavioral deficits observed do not simply reflect a lack of amidated peptides.

Original languageEnglish (US)
Article numbere28679
JournalPloS one
Volume6
Issue number12
DOIs
StatePublished - Dec 16 2011

Fingerprint

Mixed Function Oxygenases
Copper
copper
peptides
Peptides
lyases
mice
Reaction intermediates
Lyases
Glycine
vertebrates
Hydra
Vertebrates
endocrine system
neuropeptides
peptidylglycine monooxygenase
nervous system
active sites
liquid chromatography
Amyloid Precursor Protein Secretases

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Yin, P., Bousquet-Moore, D., Annangudi, S. P., Southey, B. R., Mains, R. E., Eipper, B. A., & Sweedler, J. V. (2011). Probing the production of amidated peptides following genetic and dietary copper manipulations. PloS one, 6(12), [e28679]. https://doi.org/10.1371/journal.pone.0028679

Probing the production of amidated peptides following genetic and dietary copper manipulations. / Yin, Ping; Bousquet-Moore, Danielle; Annangudi, Suresh P.; Southey, Bruce R.; Mains, Richard E.; Eipper, Betty A.; Sweedler, Jonathan V.

In: PloS one, Vol. 6, No. 12, e28679, 16.12.2011.

Research output: Contribution to journalArticle

Yin, P, Bousquet-Moore, D, Annangudi, SP, Southey, BR, Mains, RE, Eipper, BA & Sweedler, JV 2011, 'Probing the production of amidated peptides following genetic and dietary copper manipulations', PloS one, vol. 6, no. 12, e28679. https://doi.org/10.1371/journal.pone.0028679
Yin P, Bousquet-Moore D, Annangudi SP, Southey BR, Mains RE, Eipper BA et al. Probing the production of amidated peptides following genetic and dietary copper manipulations. PloS one. 2011 Dec 16;6(12). e28679. https://doi.org/10.1371/journal.pone.0028679
Yin, Ping ; Bousquet-Moore, Danielle ; Annangudi, Suresh P. ; Southey, Bruce R. ; Mains, Richard E. ; Eipper, Betty A. ; Sweedler, Jonathan V. / Probing the production of amidated peptides following genetic and dietary copper manipulations. In: PloS one. 2011 ; Vol. 6, No. 12.
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