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Analysis of the γ-secretase interactome and validation of its association with tetraspanin-enriched microdomains

Nature Cell Biology volume 11, pages 13401346 (2009) | Download Citation


γ-Secretase, an aspartyl protease that belongs to the iCLiPs (intramembrane cleaving proteases) family, is a multiprotein complex that consists of presenilin (PS), nicastrin (NCT), Aph-1 and Pen-2 (ref. 1). It is responsible for generation of the β-amyloid peptide (Aβ), the primary component of senile plaques in the brains of patients with Alzheimer's disease. Although the four components are necessary and sufficient for γ-secretase activity2,3,4, additional proteins are possibly involved in its regulation. Consequently, we purified proteins associated with the active γ-secretase complex from reconstituted PS-deficient fibroblasts, using tandem affinity purification (TAP)5 and identified a series of proteins that transiently interact with the γ-secretase complex and are probably involved in complex maturation, membrane trafficking and, importantly, the tetraspanin web. Tetraspanins form detergent-resistant microdomains in the cell membrane and regulate cell adhesion, cell signalling and proteolysis6,7. Association of the γ-secretase complex with tetraspanin-enriched microdomains provides an explanation for the previously documented localization of γ-secretase to raft-like domains8. Thus, these studies suggest that maintenance of the integrity of tetraspanin microdomains contributes to the refinement of proteolytic activity of the γ-secretase complex.

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We thank T. E. Golde (Mayo Clinic, FL, US) for SPPL3 cDNA, S. Aerts (VIB), E. Karran and the bioinformatics team of Elli-Lilly for helpful discussion and A. Thathiah for critical reading of the manuscript. This work was supported by a Pioneer award from the Alzheimer's Association, the Fund for Scientific Research, Flanders; Katholieke Universiteit Leuven (GOA); Federal Office for Scientific Affairs (IUAP P6/58) and a Methusalem grant from the Flemisch government. The laboratory in Ghent is supported by research grants from the Fund for Scientific Research – Flanders (Belgium), the Concerted Research Actions (GOA) from the Ghent University, the Inter University Attraction Poles (IUAP06) and the European Union Interaction Proteome (6th Framework Program). L.B. is fellow of the FWO (Fonds voor Wetenschappelijk Onderzoek) and T.W. was fellow of the Canon Foundation in Europe and Marie Curie program of the EC.

Author information


  1. Center for Human Genetics, K.U.Leuven, Herestraat 49, B-3000 Leuven, Belgium.

    • Tomoko Wakabayashi
    • , Katleen Craessaerts
    • , Leen Bammens
    • , Mostafa Bentahir
    •  & Bart De Strooper
  2. Department for molecular and developmental genetics, VIB, Flanders Institute for Biotechnology, Leuven, Belgium.

    • Tomoko Wakabayashi
    • , Katleen Craessaerts
    • , Leen Bammens
    • , Mostafa Bentahir
    •  & Bart De Strooper
  3. Laboratory of Medicinal Chemistry, K.U.Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium.

    • Filip Borgions
    •  & Piet Herdewijn
  4. Department of Medical Protein Research, VIB, B-9000 Ghent, Belgium.

    • An Staes
    • , Evy Timmerman
    • , Joël Vandekerckhove
    •  & Kris Gevaert
  5. Department of Biochemistry, Ghent University, B-9000 Ghent, Belgium.

    • An Staes
    • , Evy Timmerman
    • , Joël Vandekerckhove
    •  & Kris Gevaert
  6. Inserm, U602, Villejuif, France.

    • Eric Rubinstein
    •  & Claude Boucheix
  7. Université Paris 11, Institut André Lwoff, Villejuif, France.

    • Eric Rubinstein
    •  & Claude Boucheix


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T.W. designed the concept and performed most of the experiments, interpreted the data and helped write the manuscript. K.C. and L.B. performed experiments, interpreted the data and helped write the manuscript. M.B. performed initial experiments, generated cell lines, performed initial purifications of TAP-tagged proteins and helped write the manuscript. F.B. and P.H. created the γ-secretase inhibitor column, participated in discussions and helped write the manuscript. A.S. and E.T. performed LC-MS/MS experiments and analysed the results. J.V. and K.G. performed LC-MS/MS experiments and interpreted these results, supervised this part of the research and helped write the manuscript. E.C. and C.B. participated in discussions, provided advice on tetraspanin experiments and tools, and helped write the manuscript. B.D.S. designed the concept and experiments, supervised the research, coordinated experiments, interpreted the results and helped write the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Bart De Strooper.

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