ATP-binding cassette (ABC) transporters are essential molecular machines whose conformational dynamics have largely been inferred from ensemble-averaged measurements. Resolving dynamic heterogeneity and transient intermediates, however, requires single-molecule approaches. Here, we use single-molecule Förster resonance energy transfer (smFRET) to resolve ATP-driven conformational dynamics of the heterodimeric type IV ABC transporter TmrAB, a functional homolog of the human antigen transporter TAP, at the level of individual molecules. Fluorophores positioned at the nucleotide-binding domains and periplasmic gate were validated by accessible-volume simulations, fluorescence lifetimes, and ensemble FRET, demonstrating that these reporters reliably track conformational transitions. Single-molecule analysis distinguishes ATP-free and ATP-bound states and quantifies ATP-dependent population shifts from nucleotide-free to physiological ATP concentrations. Kinetic analysis further reveals an unexpectedly long ATP-bound dwell time of ~300 ms. Using complementary stabilization strategies, we directly resolve a previously hidden outward-facing open state that is kinetically masked under turnover conditions. These results provide the first single-molecule characterization of TmrAB and establish a quantitative single-molecule framework for dissecting ATP-coupled conformational dynamics in heterodimeric ABC transporters.