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

FEBS Letters

Minakata, Hiroyuki¹  Tsutsui, Kazuyoshi²  Iwakoshi, Eiko¹  Ukena, Kazuyoshi² 

[1] Suntory Institute for Bioorganic Research, Shimamoto, Mishima, Osaka 618-8503, Japan;Laboratory of Brain Science, Faculty of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima 739-8521, Japan

关键词: RFamide-related peptide;    Hypothalamus;    Immunoaffinity chromatography;    Mass spectrometry;    Rat;   

DOI  :  10.1016/S0014-5793(02)02275-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown