Quantitation of neuropeptides in Cpefat/Cpefat mice using differential isotopic tags and mass spectrometry

Fa yun Che, Lloyd D. Fricker

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Neuroendocrine peptides play important roles as intercellular messengers. We previously developed a technique to isolate and identify a large number of neuroendocrine peptides from Cpefat/Cpefat mice (Che, F.; et al. Proc. Natl. Acad. Sci. U.S.A. 2001, 98, 9971-6); these mice lack carboxypeptidase E activity and this defect causes an accumulation of neuropeptide intermediates that contain C-terminal Lys or Arg residues (Naggert, J. K.; et al. Nat. Genet. 1995, 10, 135-42). In the present study, we have developed a differential isotopic-labeling technique that can be used to quantitate changes in neuropeptide levels in Cpefat/Cpefat mouse tissues. Samples are treated with either the H6 or the D6 form of acetic anhydride, peptides that contain C-terminal basic amino acids are isolated by affinity chromatography on anhydrotrypsin agarose, and the isolated peptides are analyzed by mass spectrometry. Measurement of the regulation of pituitary peptides in response to dehydration showed a decrease in vasopressin. The general method described in this report should be widely applicable to a large number of neuroendocrine peptides, known and novel, in a variety of regulatory paradigms.

Original languageEnglish (US)
Pages (from-to)3190-3198
Number of pages9
JournalAnalytical Chemistry
Volume74
Issue number13
DOIs
StatePublished - Jul 1 2002

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Neuropeptides
Mass spectrometry
Peptides
Carboxypeptidase H
Affinity chromatography
Basic Amino Acids
Vasopressins
Dehydration
Sepharose
Labeling
Tissue
Defects

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Quantitation of neuropeptides in Cpefat/Cpefat mice using differential isotopic tags and mass spectrometry. / Che, Fa yun; Fricker, Lloyd D.

In: Analytical Chemistry, Vol. 74, No. 13, 01.07.2002, p. 3190-3198.

Research output: Contribution to journalArticle

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