Distribution of neuroendocrine regulatory peptide-1 and -2, and proteolytic processing of their precursor VGF protein in the rat.
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.
Department of Molecular Pharmacology, National Cerebral and Cardiovascular Center Research Institute, Suita, Osaka, Japan.
Emi
E
Mishiro-Sato
Kazuki
K
Sasaki
Takashi
T
Matsuo
Haruaki
H
Kageyama
Hideki
H
Yamaguchi
Yukari
Y
Date
Masako
M
Matsubara
Takehiro
T
Ishizu
Kumiko
K
Yoshizawa-Kumagaye
Yoshinori
Y
Satomi
Toshifumi
T
Takao
Seiji
S
Shioda
Masamitsu
M
Nakazato
Naoto
N
Minamino
D000818:Animals; D001923:Brain Chemistry / Q000502:physiology; D041981:Gastrointestinal Tract / Q000166:cytology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D009479:Neuropeptides / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010902:Pituitary Gland / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D014018:Tissue Distribution / Q000502:physiology
D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D010455:Peptides; D011498:Protein Precursors; C062040:Vgf protein, rat; C523894:neuroendocrine regulatory peptide-1, rat; C523896:neuroendocrine regulatory peptide-2, rat