Ectodomain shedding of neuroglycan C, a brain-specific chondroitin sulfate proteoglycan, by TIMP-2- and TIMP-3-sensitive proteolysis.
Neuroglycan C (NGC) is a transmembrane-type of chondroitin sulfate proteoglycan with an epidermal growth factor (EGF)-like module that is exclusively expressed in the CNS. Because ectodomain shedding is a common processing step for many transmembrane proteins, we examined whether NGC was subjected to proteolytic cleavage. Western blotting demonstrated the occurrence of a soluble form of NGC with a 75 kDa core glycoprotein in the soluble fraction of the young rat cerebrum. In contrast, full-length NGC with a 120 kDa core glycoprotein and its cytoplasmic fragment with a molecular size of 35 kDa could be detected in the membrane fraction. The soluble form of NGC was also detectable in culture media of fetal rat neurons, and the full-length form existed in cell layers. The amount of the soluble form in culture media was decreased by adding a physiological protease inhibitor such as a tissue inhibitor of metalloproteinase (TIMP)-2 or TIMP-3, but not by adding TIMP-1. Both EGF-like and neurite outgrowth-promoting activity of the NGC ectodomain may be regulated by this proteolytic processing.
Shuo
Takuya
T
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Aono
Sachiko
S
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Nakanishi
Keiko
K
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Tokita
Yoshihito
Y
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Kuroda
Yoshiyuki
Y
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Ida
Michiru
M
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Matsui
Fumiko
F
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Maruyama
Hiroyo
H
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Kaji
Toshiyuki
T
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
Oohira
Atsuhiko
A
Department of Perinatology and Neuroglycoscience, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Aichi, JapanDepartment of Neurochemistry, Nagoya University Graduate School of Medicine, Nagoya, JapanDepartment of Environmental Health, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, Japan.
D000818:Animals; D000831:Animals, Newborn; D000906:Antibodies / Q000494:pharmacology; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D004151:Dipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006877:Hydroxamic Acids / Q000494:pharmacology; D047468:Immunoprecipitation / Q000379:methods; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology*; D011509:Proteoglycans / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000037:antagonists & inhibitors*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000037:antagonists & inhibitors*
D000906:Antibodies; C485738:Cspg5 protein, rat; D004151:Dipeptides; D006877:Hydroxamic Acids; D008565:Membrane Proteins; C092152:N-((2-(hydroxyaminocarbonyl)methyl)-4-methylpentanoyl)-3-(2'-naphthyl)alanylalanine, 2-aminoethylamide; D011480:Protease Inhibitors; D011509:Proteoglycans; D019717:Tissue Inhibitor of Metalloproteinase-3; D019716:Tissue Inhibitor of Metalloproteinase-2