This study investigates the mechanism of human PINK1 activation at the Translocase of Outer Membrane (TOM) complex in a reconstituted system. The authors co-expressed human PINK1 and all seven TOM subunits in *Saccharomyces cerevisiae* to assess the role of each TOM subunit in PINK1 activation. They found that co-expression of wild-type human PINK1 with all TOM subunits was sufficient for PINK1 activation, and identified TOM20 and TOM70 as essential receptor subunits for optimal activation. Structural modelling using AlphaFold revealed that the N-terminal and C-terminal extensions (NTE and CTE) of PINK1 interact with the cytosolic domains of TOM20 and TOM70, respectively. Mutagenesis studies confirmed the importance of these interactions. The findings provide new insights into the regulation of PINK1 activation and could aid in the development of small molecule activators for therapeutic strategies against Parkinson's disease (PD).This study investigates the mechanism of human PINK1 activation at the Translocase of Outer Membrane (TOM) complex in a reconstituted system. The authors co-expressed human PINK1 and all seven TOM subunits in *Saccharomyces cerevisiae* to assess the role of each TOM subunit in PINK1 activation. They found that co-expression of wild-type human PINK1 with all TOM subunits was sufficient for PINK1 activation, and identified TOM20 and TOM70 as essential receptor subunits for optimal activation. Structural modelling using AlphaFold revealed that the N-terminal and C-terminal extensions (NTE and CTE) of PINK1 interact with the cytosolic domains of TOM20 and TOM70, respectively. Mutagenesis studies confirmed the importance of these interactions. The findings provide new insights into the regulation of PINK1 activation and could aid in the development of small molecule activators for therapeutic strategies against Parkinson's disease (PD).