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Novel N-terminal Cleavage of APP Precludes Aβ Generation in ACAT-Defective AC29 Cells

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Abstract

A common pathogenic event that occurs in all forms of Alzheimer’s disease is the progressive accumulation of amyloid β-peptide (Aβ) in brain regions responsible for higher cognitive functions. Inhibition of acyl-coenzyme A: cholesterol acyltransferase (ACAT), which generates intracellular cholesteryl esters from free cholesterol and fatty acids, reduces the biogenesis of the Aβ from the amyloid precursor protein (APP). Here we have used AC29 cells, defective in ACAT activity, to show that ACAT activity steers APP either toward or away from a novel proteolytic pathway that replaces both α and the amyloidogenic β cleavages of APP. This alternative pathway involves a novel cleavage of APP holoprotein at Glu281, which correlates with reduced ACAT activity and Aβ generation in AC29 cells. This sterol-dependent cleavage of APP occurs in the endosomal compartment after internalization of cell surface APP. The resulting novel C-terminal fragment APP-C470 is destined to proteasomal degradation limiting the availability of APP for the Aβ generating system. The proportion of APP molecules that are directed to the novel cleavage pathway is regulated by the ratio of free cholesterol and cholesteryl esters in cells. These results suggest that subcellular cholesterol distribution may be an important regulator of the cellular fate of APP holoprotein and that there may exist several competing proteolytic systems responsible for APP processing within the endosomal compartment.

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Acknowledgements

We thank T.Y. Chang and C.C.Y. Chang (Dartmouth Medical School, Hanover, NH, USA) for the gift of cholesterol mutant cell lines, J.H. Harwood (Pfizer, Groton, CT, USA) for providing us with the ACAT inhibitor CP-113,818, and D.J. Selkoe and W.Xia (Center for Neurological Diseases, Brigham and Women’s Hospital, Boston, MA, USA) for Aβ determinations. This work was supported by grants from the NIH–NINDS (D.M.K.), the Alzheimer’s Association (L.P.), Helsingin Sanomat Centennial Foundation (H.J.H.) and Maud Kuistila Foundation (H.J.H.).

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  1. Laura A. MacKenzie Ingano

    Present address: Merck Research Laboratories, Boston, MA, 02115, USA

  2. Luigi Puglielli

    Present address: Department of Medicine, University of Wisconsin-Madison, and Geriatric Research Education Clinical Center, VA Medical Center, Madison, WI, 53705, USA

Authors and Affiliations

  1. Neurobiology of Disease Laboratory, Genetics, and Aging Research Unit, MassGeneral Institute for Neurodegenerative Disease (MIND) and Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Charlestown, MA, 02129, USA

    Henri J. Huttunen, Luigi Puglielli, Blake C. Ellis, Laura A. MacKenzie Ingano & Dora M. Kovacs

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  1. Henri J. Huttunen
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  2. Luigi Puglielli
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Correspondence to Dora M. Kovacs.

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Huttunen, H.J., Puglielli, L., Ellis, B.C. et al. Novel N-terminal Cleavage of APP Precludes Aβ Generation in ACAT-Defective AC29 Cells. J Mol Neurosci 37, 6–15 (2009). https://doi.org/10.1007/s12031-008-9088-0

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