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著作: Dunys Julie/[瓦井 俊孝]/Wilk Sherwin/St George-Hyslop Peter/Alves da Costa Cristine/Checler Frédéric/Catabolism of endogenous and overexpressed APH1a and PEN2: evidence for artifactual involvement of the proteasome in the degradation of overexpressed proteins./[The Biochemical Journal]

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EID
347445
EOID
930458
Map
0
LastModified
2018年11月15日(木) 19:33:37
Operator
瓦井 俊孝
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TRUE
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Owner
瓦井 俊孝
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種別 必須 学術論文(審査論文)
言語 必須 英語
招待 推奨
審査 推奨
カテゴリ 推奨
共著種別 推奨
学究種別 推奨
組織 推奨
著者 必須
  1. (英) Dunys Julie
    役割 任意
    貢献度 任意
    学籍番号 推奨
  2. 瓦井 俊孝
    役割 任意
    貢献度 任意
    学籍番号 推奨
  3. (英) Wilk Sherwin
    役割 任意
    貢献度 任意
    学籍番号 推奨
  4. (英) St George-Hyslop Peter
    役割 任意
    貢献度 任意
    学籍番号 推奨
  5. (英) Alves da Costa Cristine
    役割 任意
    貢献度 任意
    学籍番号 推奨
  6. (英) Checler Frédéric
    役割 任意
    貢献度 任意
    学籍番号 推奨
題名 必須

(英) Catabolism of endogenous and overexpressed APH1a and PEN2: evidence for artifactual involvement of the proteasome in the degradation of overexpressed proteins.

副題 任意
要約 任意

(英) PS (presenilin)-dependent gamma-secretase occurs as a high-molecular-mass complex composed of either PS1 or PS2 associated with Nct (nicastrin), PEN2 (presenilin enhancer 2 homologue) and APH1 (anterior pharynx defective 1 homologue). Numerous reports have documented the very complicated physical and functional cross-talk between these proteins that ultimately governs the biological activity of the gamma-secretase, but very few studies examined the fate of the components of the complex. We show that, in both HEK-293 cells and the TSM1 neuronal cell line, the immunoreactivities of overexpressed myc-tagged-APH1a and -PEN2 were enhanced by the proteasome inhibitors ZIE and lactacystin, whereas a broad range of protease inhibitors had no effect. By contrast, proteasome inhibitors were totally unable to affect the cellular expression of endogenous APH1aL and PEN2 in HEK-293 cells, TSM1 and primary cultured cortical neurons. To explain this apparent discrepancy, we examined the degradation of myc-tagged-APH1a and -PEN2, in vitro, by cell extracts containing endogenous proteasome and by purified 20S proteasome. Strikingly, myc-tagged-APH1a and -PEN2 resist proteolysis by endogenous proteasome and purified 20S proteasome. We also show that endogenous PEN2 expression was drastically higher in wild-type than in PS- and Nct-deficient fibroblasts and was enhanced by proteasome inhibitors only in the two deficient cell systems. However, here again, purified 20S proteasome appeared unable to cleave endogenous PEN2 present in PS-deficient fibroblasts. The levels of endogenous APH1aL-like immunoreactivity were not modified by proteasome inhibitors and were unaffected by PS deficiency. Altogether, our results indicate that endogenous PEN2 and APH1aL do not undergo proteasomal degradation under physiological conditions in HEK-293 cells, TSM1 cells and fibroblasts and that the clearance of PEN2 in PS- and Nct-deficient fibroblasts is not mediated by 20S proteasome. Whether the 26S proteasome participates to PEN2 proteolysis in deficient fibroblasts remains to be established.

キーワード 推奨
  1. (英) Animals
  2. (英) Cattle
  3. (英) Cell Line
  4. (英) Endopeptidases
  5. (英) Fibroblasts
  6. (英) Gene Deletion
  7. (英) Gene Expression
  8. (英) Humans
  9. (英) Membrane Proteins
  10. (英) Mice
  11. (英) Neurons
  12. (英) Proteasome Endopeptidase Complex
  13. (英) Proteasome Inhibitors
  14. (英) Proteins
  15. (英) Receptors, Notch
  16. (英) Transfection
発行所 推奨
誌名 必須 The Biochemical Journal([The Biochemical Society])
(pISSN: 0264-6021, eISSN: 1470-8728)
ISSN 任意 1470-8728
ISSN: 0264-6021 (pISSN: 0264-6021, eISSN: 1470-8728)
Title: The Biochemical journal
Title(ISO): Biochem J
Publisher: Portland Press, Ltd.
 (NLM Catalog  (Scopus  (CrossRef (Scopus information is found. [need login])
必須 394
必須 Pt 2
必須 501 509
都市 任意
年月日 必須 2006年 3月 1日
URL 任意
DOI 任意 10.1042/BJ20051197    (→Scopusで検索)
PMID 任意 16302845    (→Scopusで検索)
NAID 任意
WOS 任意
Scopus 任意
評価値 任意
被引用数 任意
指導教員 推奨
備考 任意
  1. (英) PublicationType: Journal Article

  2. (英) PublicationType: Research Support, Non-U.S. Gov't