Alteration of mitochondria-associated membranes (MAMs) has been proposed to contribute to the pathogenesis of Alzheimer’s disease (AD). We studied herein the subcellular distribution, the processing, and the protein interactome of the amyloid-protein precursor (APP) and its proteolytic products in MAMs. We reveal that APP and its catabolites are present in MAMs in cellular models overexpressing wild type APP or APP harboring the double Swedish or London familial AD mutations, and in brains of transgenic mice model of AD. Furthermore, we evidenced that both-andsecretases are present and harbor APP processing activities in MAMs. Interestingly, cells overexpressing APPswe show increased ER-mitochondria contact sites. We also document increased neutral lipid accumulation linked to A production and reversed by inhibiting-or-secretases. Using a proteomic approach, we show that APP and its catabolites interact with key proteins of MAMs controlling mitochondria and ER functions. These data highlight the role of APP processing and proteomic interactome in MAMs deregulation taking place in AD.