A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry

Kazuyoshi Ukena, Eiko Iwakoshi, Hiroyuki Minakata, Kazuyoshi Tsutsui

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

Recently, cDNAs encoding novel RFamide-related peptides (RFRPs) have been reported in the mammalian brains by a gene database search and the deduced RFRPs have been suggested to participate in neuroendocrine and pain mechanisms in the rat. Two peptides have been predicted to be encoded in the cDNA of rodent RFRPs. To assess precise functions of rodent RFRPs in the brain, in the present study we identified a naturally occurring RFRP in the rat hypothalamus by immunoaffinity purification combined with mass spectrometry (MS). The affinity chromatography showed that the rat hypothalamus contained RFRP-like immunoreactivity. The immunoreactive material was analyzed by a nanoflow electrospray ionization time-of-flight MS followed by tandem MS analysis. The mass peak corresponding to octadecapeptide was detected at 1010.54 m/z ([M+2H]2+) and its sequence, ANMEAGTMSHFPSLPQRF-NH2, was revealed by the fragmentation, showing a mature form encoded in the cDNA sequence of RFRPs. The identified endogenous RFRP will aid not only in defining its physiological roles but also facilitate the development of its agonists and antagonists in the rodent brain.

Original languageEnglish
Pages (from-to)255-258
Number of pages4
JournalFEBS Letters
Volume512
Issue number1-3
DOIs
Publication statusPublished - 2002 Feb 13
Externally publishedYes

Keywords

  • Hypothalamus
  • Immunoaffinity chromatography
  • Mass spectrometry
  • Rat
  • RFamide-related peptide

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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