Distribution of neuroendocrine regulatory peptide-1 and -2, and proteolytic processing of their precursor VGF protein in the rat

Authors: Mishiro-Sato, Emi; Sasaki, Kazuki¹; Matsuo, Takashi²; Kageyama, Haruaki³; Yamaguchi, Hideki²; Date, Yukari⁴; Matsubara, Masako¹; Ishizu, Takehiro⁵; Yoshizawa-Kumagaye, Kumiko⁵; Satomi, Yoshinori; Takao, Toshifumi⁶; Shioda, Seiji³; Nakazato, Masamitsu²; Minamino, Naoto¹

Source: Journal of Neurochemistry, Volume 114, Number 4, August 2010 , pp. 1097-1106(10)

Publisher: Wiley-Blackwell

Abstract:

J. Neurochem. (2010) 114, 1097-1106. Abstract

Neuroendocrine regulatory peptide (NERP)-1 and NERP-2 are biologically active peptides recently discovered by peptidomic analysis. NERPs are processed out from the 594-residue VGF protein which contains many prohormone convertase cleavage motifs. VGF-deficient mice exhibit a hypermetabolic and infertile phenotype, for which VGF protein-derived peptides including NERPs are presumably responsible. To provide a solid basis for elucidating physiological roles of NERPs, we investigated rat VGF protein processing by chromatographic and mass spectrometric analysis, and immunoblotting, using antibodies against NERPs and the VGF protein C-terminus (VGF-C). Cellular and tissue distribution of immunoreactive (ir) NERPs were also analyzed in the rat. Both ir-NERP-1 and ir-NERP-2, which occur abundantly in the CNS and pituitary, moderately in the gastrointestinal (GI) tract, were mainly localized in neuronal structures. Major endogenous forms of ir-NERPs in the brain and GI tract were identified as NERP-1, NERP-2, and big NERP-2 (NERP-1 + NERP-2), with NERP-1 and big NERP-2 being predominant. Regarding ir-VGF-C peptides, VGF[588-617], VGF[556-617], and VGF[509-617] were found to be major forms. Immunoblotting with the NERP-2 and VGF-C antibodies revealed processing intermediates of 10-37 kDa. Taken together, we deduce that VGF protein is primarily cleaved at 10 sites through the processing pathway common to the brain and GI tract.

Keywords: immunohistochemistry; multi-peptide hormone precursor; neuroendocrine regulatory peptide; peptidomics; proteolytic processing; VGF protein

Document Type: Research article

DOI: http://dx.doi.org/10.1111/j.1471-4159.2010.06827.x

Affiliations: 1: Department of Molecular Pharmacology, National Cerebral and Cardiovascular Center Research Institute, Suita, Osaka, Japan 2: Division of Neurology, Respirology, Endocrinology and Metabolism, Department of Internal Medicine, Miyazaki Medical College, University of Miyazaki, Kiyotake, Miyazaki, Japan 3: Department of Anatomy, Showa University School of Medicine, Shinagawa, Tokyo, Japan 4: Frontier Science Research Center, University of Miyazaki, Kiyotake, Miyazaki, Japan 5: Saito Research Center, Peptide Institute, Inc., Ibaraki, Osaka, Japan 6: Institute for Protein Research, Osaka University, Suita, Osaka, Japan

Publication date: 2010-08-01

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• In this Subject: Anatomy & Physiology
• By this author: Mishiro-Sato, Emi ;  Sasaki, Kazuki ;  Matsuo, Takashi ;  Kageyama, Haruaki ;  Yamaguchi, Hideki ;  Date, Yukari ;  Matsubara, Masako ;  Ishizu, Takehiro ;  Yoshizawa-Kumagaye, Kumiko ;  Satomi, Yoshinori ;  Takao, Toshifumi ;  Shioda, Seiji ;  Nakazato, Masamitsu ;  Minamino, Naoto

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