Purification and cloning of amyloid precursor protein β-secretase from human brain (original) (raw)

• Letter
• Published: 02 December 1999
• John P. Anderson1,
• Robin Barbour1,
• Guriqbal S. Basi1,
• Russell Caccavello1,
• David Davis1,
• Minhtam Doan1,
• Harry F. Dovey1,
• Normand Frigon1,
• Jin Hong1,
• Kirsten Jacobson-Croak1,
• Nancy Jewett1,
• Pamela Keim1,
• Jeroen Knops1,
• Ivan Lieberburg1,
• Michael Power1,
• Hua Tan1,
• Gwen Tatsuno1,
• Jay Tung1,
• Dale Schenk1,
• Peter Seubert1,
• Susanna M. Suomensaari1,
• Shuwen Wang1,
• Donald Walker1,
• Jun Zhao1,
• Lisa McConlogue1 &
• …
• Varghese John1

Nature volume 402, pages 537–540 (1999)Cite this article

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Abstract

Proteolytic processing of the amyloid precursor protein (APP) generates amyloid β (Aβ) peptide, which is thought to be causal for the pathology and subsequent cognitive decline in Alzheimer's disease. Cleavage by β-secretase at the amino terminus of the Aβ peptide sequence, between residues 671 and 672 of APP, leads to the generation and extracellular release of β-cleaved soluble APP1, and a corresponding cell-associated carboxy-terminal fragment. Cleavage of the _C_-terminal fragment by γ-secretase(s) leads to the formation of Aβ. The pathogenic mutation K670M671 → N670L671 at the β-secretase cleavage site in APP2, which was discovered in a Swedish family with familial Alzheimer's disease, leads to increased β-secretase cleavage of the mutant substrate3. Here we describe a membrane-bound enzyme activity that cleaves full-length APP at the β-secretase cleavage site, and find it to be the predominant β-cleavage activity in human brain. We have purified this enzyme activity to homogeneity from human brain using a new substrate analogue inhibitor of the enzyme activity, and show that the purified enzyme has all the properties predicted for β-secretase. Cloning and expression of the enzyme reveals that human brain β-secretase is a new membrane-bound aspartic proteinase.

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References

1. Seubert,P. et al. Secretion of beta-amyloid precursor protein cleaved at the amino terminus of the beta-amyloid peptide. Nature 361, 260–263 (1993).
   Article ADS CAS Google Scholar
2. Mullan,M. et al. A pathogenic mutation for probably Alzheimer's disease in the APP gene at the N-terminus of beta-amyloid. Nature Genet. 1, 345–347 (1992).
   Article CAS Google Scholar
3. Citron,M. et al. Mutation of the beta-amyloid precursor protein in familial Alzheimer's disease increases beta-protein secretion. Nature 360, 672–674 (1992).
   Article ADS CAS Google Scholar
4. Knops,J. et al. Cell-type and amyloid protein-type specific inhibition of Ab release by bafilomycin A1, a selective inhibitor of vacuolar ATPases. J. Biol. Chem. 270, 2419–2422 (1995).
   Article CAS Google Scholar
5. Valdes,F., Munoz,C., Feria-Velasco,A. & Orrego,F. Subcellular distribution of rat brain cortex high-affinity, sodium-dependent, glycine transport sites. Brain Res. 122, 95–112 (1977).
   Article CAS Google Scholar
6. Narutaki,S., Dunn,B. M. & Oda,K. Subsite preference of pepstatin-insensitive carboxyl proteinases from bacteria. J. Biochem. (Tokyo) 125, 75–81 (1999).
   Article CAS Google Scholar
7. Schechter,I. & Berger,A. On the size of the active site in proteinases. I. Papain. Biochem. Biophys. Res. Commun. 27, 157–162 (1967).
   Article CAS Google Scholar
8. Rich,D. H. in Proteinase Inhibitors (eds Barrett, A. J. & Salvesen, G.) (Elsevier Science, Amsterdam, 1986).
   Google Scholar
9. Johnson-Wood,K. et al. Amyloid precursor protein processing and Aβ42 deposition in a transgenic mouse model of Alzheimer disease. Proc. Natl Acad. Sci. USA 94, 1550–1555 (1997).
   Article ADS CAS Google Scholar
10. Vassar,R. et al. β-secretase cleavage of the Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. Science 286, 735–741 (1999).
    Article CAS Google Scholar
11. Lin,J. H., Ostovic,D. & Vacca,J. P. The integration of medicinal chemistry, drug metabolism, and pharmaceutical research and development. The story of Crixivan, an HIV proteinase inhibitor. Pharm. Biotechnol. 11, 233–255 (1998).
    Article CAS Google Scholar
12. Knops,J. et al. Isolation of Baculovirus-derived secreted and full-length β-amyloid precursor protein. J. Biol. Chem. 266, 7285–7290 (1991).
    CAS PubMed Google Scholar

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Authors and Affiliations

1. Elan Pharmaceuticals, 800 Gateway Boulevard, South San Francisco, 94080, California, USA
   Sukanto Sinha, John P. Anderson, Robin Barbour, Guriqbal S. Basi, Russell Caccavello, David Davis, Minhtam Doan, Harry F. Dovey, Normand Frigon, Jin Hong, Kirsten Jacobson-Croak, Nancy Jewett, Pamela Keim, Jeroen Knops, Ivan Lieberburg, Michael Power, Hua Tan, Gwen Tatsuno, Jay Tung, Dale Schenk, Peter Seubert, Susanna M. Suomensaari, Shuwen Wang, Donald Walker, Jun Zhao, Lisa McConlogue & Varghese John

Authors

1. Sukanto Sinha
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2. John P. Anderson
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3. Robin Barbour
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4. Guriqbal S. Basi
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5. Russell Caccavello
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6. David Davis
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7. Minhtam Doan
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8. Harry F. Dovey
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9. Normand Frigon
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10. Jin Hong
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11. Kirsten Jacobson-Croak
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12. Nancy Jewett
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13. Pamela Keim
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14. Jeroen Knops
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15. Ivan Lieberburg
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16. Michael Power
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17. Hua Tan
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18. Gwen Tatsuno
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19. Jay Tung
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20. Dale Schenk
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21. Peter Seubert
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22. Susanna M. Suomensaari
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23. Shuwen Wang
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24. Donald Walker
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25. Jun Zhao
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26. Lisa McConlogue
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27. Varghese John
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Corresponding author

Correspondence toSukanto Sinha.

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Sinha, S., Anderson, J., Barbour, R. et al. Purification and cloning of amyloid precursor protein β-secretase from human brain.Nature 402, 537–540 (1999). https://doi.org/10.1038/990114

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• Received: 01 November 1999
• Accepted: 08 November 1999
• Issue Date: 02 December 1999
• DOI: https://doi.org/10.1038/990114