A novel molecular mechanism for ectodomain shedding

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 18590809
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Circulatory organs internal medicine
• Research Institution: Kyoto University
• Principal Investigator: NISHI Eiichiro Kyoto University, Department of Cardiovascular Medicine, Assistant Professor (30362528)
• Project Period (FY): 2006 – 2007
• Keywords: Ectodomain shedding / Membrane protein / Growth factors / Cytokines / Metalloprteases / Nardilysin / ADAM proteases

Research Abstract

Ectodomain shedding is an irreversible posttranslational modification that releases the extracellular domain of membrane-anchored protein through proteolysis Ectodomain shedding of some cytokines and growth factors, such as TNF-alpha and EGF family members, converts the membrane-anchored pro-form to active soluble form. The generation of soluble TNF- alpha from membrane-bound TNF- alpha involves an enzyme activity denoted TNF- alpha converting enzyme (TACE). However, the mechanism that regulates TACE activity remains unclear. We have recently demonstrated that nardilysin (N-arginine dibasic convertase; NRDc), a metalloendopeptidase of M16 family, enhanced ectodomain shedding of heparin-binding epidermal growth factor-like growth factor (HB-EGF) by activation of TACE (Nishi, Hiraoka, et. al., J. Biol. Chem., 2006). NRDc binds to TACE and directly enhances the catalytic activity of TACE, which was the first description how ectodomain shedding is regulated by modulation of sheddase activi ty. We have also shown that NRDc enhances ectodomain shedding of several other substrates of TACE such as amyloid precursor protein (Hiraoka, et. al., J. Neurochem., 2007) and TNF- alpha (Hiraoka, et. al., BBRC, 2008), suggesting that NRDc is involved in the ectodomain shedding of a broad spectrum membrane proteins, and that it regulates various pathological processes through the enhancement of ectodomain shedding of those membrane proteins.
To clarify the biological roles of NRDc in vivo, we have recently generated NRDc-deficient mice by a targeted disruption of the NRDc gene locus. While homozygous mutant of NRDc-deficient mice were born at the expected ratio according to Mendel's law, they showed obvious pre- and past-natal growth retardation. Analysis by CT scan surprisingly showed that the volume of adipose tissue (corrected by body weight) in NRDc-deficient mice is much less than that in wild-type (WT) mice (white adipose tissue: 47% of WT, brown adipose tissue: 24% of WI). Furthermore, NRDc-deficient mice displayed significantly enhanced insulin sensitivity in a glucose tolerance test as compared to the control mice. These results suggested that NRDc plays essential roles in carbohydrate and lipid metabolisms in vivo.
Further studies on this proposal may provide a new insight for the biological and pathophysiological roles of ectodomain shedding in vivo.

Research Products

• [Journal Article] Ectodomain shedding of TNF-alpha is enhanced by nardilysin via activation of ADAM proteases. (2008)
  • Author(s): Hiroka Y, Yoshida K, Ohno M, Matsuoka T, Kita T, Nishi E.
  • Journal Title: Biochem. Biophs. Res. Commun. 370 Pages: 154-158
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Identification of adherens junction-associated GTPase activating proteins by the fluorescence localization-based expression cloning (2008)
  • Author(s): Matsuda M, Kobayashi Y, Masuda S, Adachi M, Watanabe T, Yamashita JK, Nishi E, Tsukita S, Furuse M.
  • Journal Title: Exp Cell Res. 314 Pages: 939-949
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Ectodomain shedding of TNF-alpha is enhanced by nardilysin via activation of ADAM proteases (2008)
  • Author(s): Hiraoka, Y., Yoshida, K., Ohno, M., Matsuoka, T., Kita, T., Nishi, E
  • Journal Title: Biochem. Biophs. Res. Commun 370 Pages: 154-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of adherens junction-associated GTPase activating proteins by the fluorescence localization-based expression cloning (2008)
  • Author(s): Matsuda, M., Kobayashi, Y., Masuda, S., Adachi, M., Watanabe, T., Yamashita, JK., Nishi, E., Tsukita, S., Furuse, M
  • Journal Title: Exp Cell Res 314 Pages: 939-49
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Platelets Are Novel Regulators of Neovascularization and Luteinization during Human Corpus Luteum Formation. (2007)
  • Author(s): Furukawa K, Fujiwara H, Sato Y, Zeng BX, Fujii H, Yoshioka S, Nishi E, Nishio T.
  • Journal Title: Endocrinology 148 Pages: 3056-3064
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Enhancement of alpha-secretase cleavage of amyloid precursor protein by a metalloendopeptidase nardilysin. (2007)
  • Author(s): Hiraoka Y, Ohno M, Yoshida K, Okawa K, Tomimoto H, Kita T, Nishi E.
  • Journal Title: J Neurochem. 102 Pages: 1595-1605
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Platelets Are Novel Regulators of Neovascularization and Luteinization during Human Corpus Luteum Formation (2007)
  • Author(s): Furukawa, K., Fujiwara, H., Sato, Y., Zeng, BX., Fujii, H., Yoshioka, S., Nishi, E., Nishio, T
  • Journal Title: Endocrinology 148 Pages: 3056-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Enhancement of alpha-secretase cleavage of amyloid precursor protein by a metalloendopeptidase nardilysin (2007)
  • Author(s): Hiraoka, Y., Ohno, M., Yoshida, K., Okawa, K., Tomimoto, H., Kita, T., Nishi, E
  • Journal Title: J Neurochem 148 Pages: 3056-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Nardilysin enhances ectodomain shedding of heparin-binding EGF-like growth factor through activation of TNF-alpha converting enzyme (2006)
  • Author(s): Nishi E, Hiraoka Y, Yoshida K, Okawa K, Kita T
  • Journal Title: J Biol Chem. 281 Pages: 31164-31172
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Nardilysin enhances ectodomain shedding of heparin-binding EGF-like growth factor through activation of TNF-alpha converting enzyme (2006)
  • Author(s): Nishi, E., Hiraoka, Y., Yoshida, K., Okawa, K., Kita, T
  • Journal Title: J Biol Chem 281 Pages: 31164-72
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A Metalloendopeptidase Nardilysin, Activator for Ectodomain Shedding, is an Essential Regulator for Energy Metabolism (2008)
  • Author(s): Eiichiro, Nishi, Hiraoka, Y., Yoshida, K., Ohno, M., Matsuoka, T., Kita, T
  • Organizer: The Mid Annual Scientific Meeting of the Japanese Circulation Society
  • Place of Presentation: Fukuoka
  • Year and Date: 20080328-30
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A Metalloendopeptidase Nardilysin, Activator for Ectodomain Shedding, is an Essential Regulator for Energy Metabolism (2008)
  • Author(s): Nishi, E, Hlraoka, Y, Yoshida, K, Ohno, M, Matsuoka, T, Kita, T
  • Organizer: The 72nd Annual Scientific Meeting of the Japanese Circulation Society
  • Place of Presentation: 福岡
  • Year and Date: 2008-03-29
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Molecular mechanism in which ectodomain shedding is enhanced (2007)
  • Author(s): Eiichiro, Nishi, Hiraoka, Y., Yoshida, K., Ohno, M., Matsuoka, T., Kita, T
  • Organizer: BMB2007
  • Place of Presentation: Yokohama
  • Year and Date: 20071211-15
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 細胞外ドメインシェディング誘導の分子機構 (2007)
  • Author(s): 西英一郎、平岡義範、吉田和弘、大野美紀子、松岡龍彦、北徹
  • Organizer: BMB2007(第30回日本分子生物学会年会、第80回日本生化学会大会 合同大会)
  • Place of Presentation: 横浜
  • Year and Date: 2007-12-15
  • Description: 「研究成果報告書概要(和文)」より