Tissue-Nonspecific Alkaline Phosphatase with an Asp289→Val Mutation Fails to Reach the Cell Surface and Undergoes Proteasome-Mediated Degradation

• Ishida Yoko

  Division of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences Removable Prosthodontics, Niigata University Graduate School of Medical and Dental Sciences

• Komaru Keiichi

  Division of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences Kitasato Junior College of Health and Hygienic Sciences

• Ito Masahiro

  Division of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences

• Amaya Yoshihiro

  Division of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences

• Kohno Shoji

  Removable Prosthodontics, Niigata University Graduate School of Medical and Dental Sciences

• Oda Kimimitsu

  Division of Biochemistry, Niigata University Graduate School of Medical and Dental Sciences Center for Transdisciplinary Research, Niigata University

書誌事項

タイトル別名

• Tissue-Nonspecific Alkaline Phosphatase with an Asp²⁸⁹→Val Mutation Fails to Reach the Cell Surface and Undergoes Proteasome-Mediated Degradation
• Tissue Nonspecific Alkaline Phosphatase with an Asp289 Val Mutation Fails to Reach the Cell Surface and Undergoes Proteasome Mediated Degradation

抄録

A missense mutation in the gene of tissue-nonspecific alkaline phosphatase, which replaces aspartic acid at position 289 with valine [TNSALP (D289V)], was reported in a lethal hypophosphatasia patient [Taillandier, A. et al. (1999) Hum. Mut. 13, 171-172]. To define the molecular defects of TNSALP (D289V), this mutant protein in transiently transfected COS-1 cells was analyzed biochemically and morphologically. TNSALP (D289V) exhibited no alkaline phosphatase activity and mainly formed a disulfide-linked high molecular mass aggregate. Cell-surface biotinylation, digestion with phosphatidylinositol-specific phospholipase C and an immunofluorescence study showed that the mutant protein failed to appear on the cell surface and was accumulated intracellularly. In agreement with this, pulse/chase experiments demonstrated that TNSALP (D289V) remained endo-β-N-acetyl- glucosaminidase H-sensitive throughout the chase and was eventually degraded, indicating that the mutant protein is unable to reach the medial-Golgi. Proteasome inhibitors strongly blocked the degradation of TNSALP (D289V), and furthermore the mutant protein was found to be ubiquitinated. Besides, another naturally occurring TNSALP with a Glu²¹⁸→Gly mutation was also found to be polyubiquitinated and degraded in the proteasome. Since the acidic amino acids at positions 218 and 289 of TNSALP are thought to be directly involved in the Ca²⁺ coordination, these results suggest the critical importance of calcium binding in post-translational folding and assembly of the TNSALP molecule.

収録刊行物

• The Journal of Biochemistry

  The Journal of Biochemistry 134 (1), 63-70, 2003

  社団法人 日本生化学会

キーワード

• alkaline phosphatase
• calcium
• hypophosphatasia
• proteasome
• ubiquitination

詳細情報

• CRID: 1390001204963934848
• NII論文ID: 130003534559
• NII論文ID: 50000881639
• NII書誌ID: AA00694073
• DOI: 10.1093/jb/mvg114
• COI: 1:CAS:528:DC%2BD3sXnslGlurk%3D
• ISSN: 17562651
• ISSN: 0021924X
• NDL書誌ID: 6640687
• PubMed: 12944372
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I6640687
• Web Site: http://academic.oup.com/jb/article-pdf/134/1/63/2447476/mvg114.pdf
• Web Site: https://search.jamas.or.jp/link/ui/2003316339
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可