A study for the sorting mechanism of proteins to Secretory Granules in Neuroendocrine cells : Emerging Concepts with Granins.

• Project/Area Number: 15500205
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Neuroscience in general
• Research Institution: Gunma University
• Principal Investigator: HOSAKA Masanori Gunma University, Institute for Molecular and Cellular Regulation, Associate Professor, 生体調節研究所, 助教授 (80311603)
• Project Period (FY): 2003 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥3,700,000 (Direct Cost: ¥3,700,000); Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000); Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
• Keywords: neuroendocrine cell / peptide-hormone / neuro-peptide / secretory granule / sorting mechanism / secretogranin III / carboxypeptidase E / cholesterol / セレクトグラニンIII / プロホルモン / 顆粒膜 / リポソーム / スフィンゴリピッド / リピッドラフト

Research Abstract

Granin-family proteins, including chromogranin A(CgA), chromogranin B, secretogranin II, secretogranin III(SgIII), and 7B2, are thought to condense peptide hormones to SGs of neuroendocrine cells. I previously showed that SgIII targets CgA to SGs in neuroendocrine cells. In this study, we investigated the sorting mechanism of SgIII to the SGs and the interaction between SgIII and carboxypeptidase E(CPE;a different candidate molecule of a sorting receptor for prohormones).
The sorting mechanism of SgIII to the SGs ; Analyses with subcellular fractionation showed that significant amounts of SgIII are bound to the secretory granule membrane(SGM). The membrane-bound SgIII could be dissociated with Triton X-100 or depleting cholesterol of the SGM. Subsequent analyses with liposome-binding assays demonstrated that SgIII binds to the cholesterol-rich liposome that mimics the composition of the SGM. Although CgA does not bind directly to the SGM-type liposome, CgA binds to the SGM-type liposome only in the presence of SgIII. Thus, these findings suggest that SgIII directly binds to cholesterol components of the SGM and targets CgA to the SGs in neuroendocrine cells.
The interaction between SgIII and CPE ; We investigated the binding of SgIII and CPE, because they frequently localized closely. They bound each other in neuroendocrine cells. In Cpefat mouse corticotropes having defective CPE, SgIII independently localized on the secretory granule membrane. Moreover, proopiomelanocortin(POMC) distributed over the entire secretory granules, and displayed a regulated secretion by secretagogues in Cpefat mouse. The Cpefat pltuitary exhibited elevated levels of SgIII and CgA, suggesting their compensation for a sorting function of CPE for POMC. Indeed, both SgIII and CgA were able to bind POMC. These data suggest that SgIII and CPE core the separate functional sorting complex by anchoring to cholesterol-rich secretory granule membranes, and POMC-derived peptides are transferred from CPE to SgIII, and further to CgA.

Research Products

• [Journal Article] Angiogenic Endothelium-specific Nestin Expression is enhanced by the First Intron of the Nestin Gene. (2004)
  • Author(s): Aihara, M., Sugawara, K., Torii, S., Hosaka M, Kurihara, H., Saito, N., Takeuchi, T.
  • Journal Title: Lab.Invest. 84 Pages: 1581-1592
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Immunocytochemical localization of secretogranin III in the endocrine pancreas of male rats. (2004)
  • Author(s): Sakai, Y., Hosaka M., Yoshinaga, A., Hira, Y., Harumi, T., Watanabe, T.
  • Journal Title: Arch Histol Cytol. 67 Pages: 57-64
  • NAID: 10012846445
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Functional localization of proprotein -convertase furin and its substrate TGFb in EGF receptor-expressing gastric chief cells. (2004)
  • Author(s): Fujisawa, T., Kamimura, H., Hosaka, M., Torii, S., Izumi, T., Kuwano, H., Takeuchi, T.
  • Journal Title: Growth Factors 22 Pages: 51-59
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Secretogranin III binds to cholesterol in the secretory granule membrane as an adapter for chromogranin A. (2004)
  • Author(s): Hosaka, M., Suda, M., Sakai, Y., Izumi, T, Watanabe, T., Takeuchi, T.
  • Journal Title: J.Biol.Chem. 30 Pages: 3627-3634
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] PTHrP induces insulin expression through activation of MAP kinase-specific phosphatase-1 that dephosphorylates c-jun N-terminal kinase in pancreatic b-cells. (2004)
  • Author(s): Zhang, B., Hosaka, M., Sawada, Y., Torii, S., Mizutani, S., Ogata, M., Izumi, T., Takeuchi, T.
  • Journal Title: Diabetes 52 Pages: 2720-2730
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Angiogenic Endothelium-specific Nestin Expression is enhanced by the First Intron of the Nestin Gene. (2004)
  • Author(s): Aihara, M., Sugawara, K., Torii, S., Hosaka.M., Kurihara, H., Saito, N., Takeuchi, T.
  • Journal Title: Lab.Invest. 84 Pages: 1581-1592
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Immunocytochemical localization of secretogranin III in the endocrine pancreas of male rats. (2004)
  • Author(s): Sakai, Y., Hosaka, M., Yoshinaga, A., Hira, Y., Harumi, T., Watanabe, T.
  • Journal Title: Arch Histol Cytol. 67 Pages: 57-64
  • NAID: 10012846445
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Functional localization of proprotein-convertase furin and its substrate TGFb in EGF receptor-expressing gastric chief cells. (2004)
  • Author(s): Fujisawa, T., Kamimura, H., Hosaka, M., Torii, S., Izumi, T., Kuwano, H., Takeuchi, T.
  • Journal Title: Growth Factors 22 Pages: 51-59
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Angiogenic enfothelium-specific nestin expression is enhanced by the first intron of the nestin gene. (2004)
  • Author(s): Aihara M, Sugawara K, Torii S, Hosaka M, Kurihara H, Saito N
  • Journal Title: Laboratory Investigation 84 Pages: 1581-1592
• [Journal Article] Immunocytochemical localization of secretogranin III in the endocrine pancreas of male rats. (2004)
  • Author(s): Sakai Y, Hosaka M, Yoshinaga A, Hira Y, Harumi T, Watanabe T
  • Journal Title: Arch.Histol.Cytol. 67 Pages: 57-64
  • NAID: 10012846445
• [Publications] Zhang, B., Hosaka, M., Sawada, Y., Torii, S., Mizutani, S., Ogata, M., Izumi, T., Takeuchi, T.: "PTHrP induces insulin expression through activation of MAP kinase-specific phosphatase-1 that dephosphorylates c-jun N-terminal kinase in pancreatic β-cells."Diabetes. 52. 2720-2730 (2003)
• [Publications] Hosaka, M., Suda, M., Sakai, Y., Izumi, T, Watanabe, T., Takeuchi, T.: "Secretogranin III binds to cholesterol in the secretory granule membrane as an adapter for chromogranin A."J.Biol.Chem. 279. 3627-3634 (2004)