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

143 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

Fingerprint

Chromatography
Mass spectrometry
Rats
Mass Spectrometry
Peptides
Rodentia
Complementary DNA
Brain
Hypothalamus
Affinity chromatography
arginylphenylalaninamide
RFamide peptide
Electrospray ionization
Tandem Mass Spectrometry
Affinity Chromatography
Purification
Databases
Genes
Pain

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry. / Ukena, Kazuyoshi; Iwakoshi, Eiko; Minakata, Hiroyuki; Tsutsui, Kazuyoshi.

In: FEBS Letters, Vol. 512, No. 1-3, 13.02.2002, p. 255-258.

Research output: Contribution to journalArticle

Ukena, Kazuyoshi ; Iwakoshi, Eiko ; Minakata, Hiroyuki ; Tsutsui, Kazuyoshi. / A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry. In: FEBS Letters. 2002 ; Vol. 512, No. 1-3. pp. 255-258.
@article{3ae898521c4b4d139425d42c3470281c,
title = "A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry",
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.",
keywords = "Hypothalamus, Immunoaffinity chromatography, Mass spectrometry, Rat, RFamide-related peptide",
author = "Kazuyoshi Ukena and Eiko Iwakoshi and Hiroyuki Minakata and Kazuyoshi Tsutsui",
year = "2002",
month = "2",
day = "13",
doi = "10.1016/S0014-5793(02)02275-5",
language = "English",
volume = "512",
pages = "255--258",
journal = "FEBS Letters",
issn = "0014-5793",
publisher = "Elsevier",
number = "1-3",

}

TY - JOUR

T1 - A novel rat hypothalamic RFamide-related peptide identified by immunoaffinity chromatography and mass spectrometry

AU - Ukena, Kazuyoshi

AU - Iwakoshi, Eiko

AU - Minakata, Hiroyuki

AU - Tsutsui, Kazuyoshi

PY - 2002/2/13

Y1 - 2002/2/13

N2 - 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.

AB - 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.

KW - Hypothalamus

KW - Immunoaffinity chromatography

KW - Mass spectrometry

KW - Rat

KW - RFamide-related peptide

UR - http://www.scopus.com/inward/record.url?scp=0037070155&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0037070155&partnerID=8YFLogxK

U2 - 10.1016/S0014-5793(02)02275-5

DO - 10.1016/S0014-5793(02)02275-5

M3 - Article

VL - 512

SP - 255

EP - 258

JO - FEBS Letters

JF - FEBS Letters

SN - 0014-5793

IS - 1-3

ER -