DDHD2 mutation was a common cause of SPG54 disease. While its mechanism of action was unclear, we aimed to comprehensively characterize the DDHD2 interactome networks in cells. To achieve this, we used affinity purification coupled with mass spectrometry (AP-MS) to define the interactome for DDHD2. We transiently transfected FLAG-DDHD2 expression plasmids into 293T cells, and then performed whole cell lysis for FLAG immunoprecipitation followed by mass spectrometry analysis (IP-MS). As a result, we identified components of the autophagy complex that were enriched in the interactome of DDHD2