Peptidomics for studying biological functions of VGF

• Project/Area Number: 25461402
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Endocrinology
• Research Institution: National Cardiovascular Center Research Institute
• Principal Investigator: Sasaki Kazuki 国立研究開発法人国立循環器病研究センター, 研究所, 室長 (80260313)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: ペプチド / 質量分析 / プロセシング / ペプチドミクス / peptides / peptidomics / insulin / mass spectrometry / 生理活性ペプチド

Outline of Final Research Achievements

VGF represents a neurosecretory protein expressed in a subset of hypothalamic neurons and endocrine cells. It has been considered a precursor to some neuropeptides because of several pairs of basic amino acid residues. This complexity has defied conventional approaches to studying the whole picture of its processing, which would provide a clue to elucidating the biological function. Alternatively we have advocated a mass spectrometry-based peptide profiling method with an aim of inspecting peptides secreted by VGF-expressing endocrine cells. Our study has revealed that the precursor undergoes limited proteolysis to more than 15 processing products, with a diverse set of post-translational modifications. One of the processed peptides proved to be an endogenous suppressor of insulin secretion. Our peptidomics approach would be extended to different proteins without relying on antibodies.

Research Products

• [Journal Article] Peptidomics for Studying Limited Proteolysis (2015)
  • Author(s): Takashi Tsuchiya, Tsukasa Osaki, Naoto Minamino, Kazuki Sasaki
  • Journal Title: Journal of Proteome Research Volume: 14 Issue: 11 Pages: 4921-4931
  • DOI: 10.1021/acs.jproteome.5b00820
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Chronic effects of neuroendocrine regulatory peptide (NERP-1 and -2) on insulin secretion and gene expression in pancreatic beta-cells. (2015)
  • Author(s): Kim JW, Rhee M, Park JH, Yamaguchi H, Sasaki K, Minamino N, Nakazato M, Song DK, Yoon KH
  • Journal Title: Biochem Biophys Res Commun Volume: 457 Issue: 2 Pages: 148-153
  • DOI: 10.1016/j.bbrc.2014.12.067
  • Peer Reviewed / Open Access
• [Journal Article] Region-specific activation of CRTC1-CREB signaling mediates long-term fear memory (2014)
  • Author(s): Nonaka M, Kim R, Fukushima H, Sasaki K, Suzuki K, Okamura M, Ishii Y, Kawashima T, Kamijo S, Takemoto-Kimura S, Okuno H, Kida S, Bito H
  • Journal Title: Neuron Volume: 84 Issue: 1 Pages: 92-106
  • DOI: 10.1016/j.neuron.2014.08.049
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Flotillin-1 regulates oncogenic signaling in neuroblastoma cells by regulating ALK membrane association. (2014)
  • Author(s): Tomiyama A, Uekita T, Kamata R, Sasaki K, Takita J, Ohira M, Nakagawara A, Kitanaka C, Mori K, Yamaguchi H, Sakai R.
  • Journal Title: Cancer Res. Volume: 74(14) Issue: 14 Pages: 3790-3801
  • DOI: 10.1158/0008-5472.can-14-0241
  • Peer Reviewed / Open Access
• [Journal Article] Neuroendocrine regulatory peptide-1 and -2 (NERPs) inhibit the excitability of magnocellular neurosecretory cells in the hypothalamus. (2014)
  • Author(s): Toshinai K, Saito T, Yamaguchi H, Sasaki K, Tsuchimochi W, Minamino N, Ueta Y, Nakazato M
  • Journal Title: Brain Res Volume: 1563 Pages: 52-60
  • DOI: 10.1016/j.brainres.2014.03.038
  • Peer Reviewed / Open Access
• [Journal Article] ペプチドミクス―内因性分泌ペプチドの一斉解析から新規生理活性ペプチドの発見へ (2014)
  • Author(s): 佐々木一樹
  • Journal Title: 医学のあゆみ Volume: 251 Pages: 1017-1021
• [Journal Article] Continuous stress promotes expression of VGF in melanotroph via suppression of dopamine (2013)
  • Author(s): Kyohei Tokizane
  • Journal Title: Molecular and Cellular Endocrinology Volume: 372 Issue: 1-2 Pages: 4956-4956
  • DOI: 10.1016/j.mce.2013.03.012
  • Peer Reviewed
• [Presentation] Utility of electron transfer dissociation mass spectrometry in global analysis of naturally occurring peptides
  • Author(s): Sasaki K, Osaki T, Minamino N
  • Organizer: 4th Asia-Pacific International Peptide Symposium
  • Place of Presentation: 大阪
• [Presentation] Peptidomic survey of peptides found in culture supernatant of cardiac fibroblasts-an expanded application to common cells having the constitutive secretory pathway
  • Author(s): Tsuchiya T, Sasaki K, Osaki T, Minamino N
  • Organizer: 4th Asia-Pacific International Peptide Symposium
  • Place of Presentation: 大阪
• [Book] Mass Spectrometry-based Peptidomics for the Discovery of Antimicrobial Peptides (2013)
  • Author(s): Tsukasa Osaki, Kazuki Sasaki and Naoto Minamino
  • Total Pages: 9
  • Publisher: Formatex Research Center
• [Remarks] ペプチドーム解析に基づいた生理活性ペプチドの探索の詳細
  • URL: http://www.ncvc.go.jp/res/divisions/pharmacology/001/a_detail.html