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 (AβPP) and its proteolytic products in MAMs. We reveal that A PP and its catabolites are present in MAMs in cellular models overexpressing wild type A PP or A PP harboring the double Swedish or Londonfamilial AD mutations, and in brains of transgenic mice model of AD. Furthermore, we evidenced that both - and -secretases are present and harbor A PP processing activities in MAMs. Interestingly, cells overexpressing APPsweshowincreased 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 A PP and its catabolites interact with key proteins of MAMs controlling mitochondria and ER functions. These data highlight the role of A PP processing and proteomic interactome in MAMs deregulation taking place in AD.