Global mapping of protein localization in vegetative Dictyostelium discoideum by subcellular spatial proteomics

Dictyostelium discoideum is a genetically tractable amoebozoan model organism widely used to study conserved eukaryotic cellular processes, yet a comprehensive map of its subcellular proteome has been lacking. Here, we combined subcellular fractionation with label-free mass spectrometry to resolve a global protein localization map for D. discoideum . We detected 6,337 proteins (~50% of the predicted proteome) and generated protein abundance profiles across 10 subcellular fractions. Using a curated marker set and a support vector machine classifier with a median cutoff, we assigned high-confidence localizations to 3,169 proteins across 17 subcellular compartments. Independent validation using sequence-based targeting predictions strongly supported the compartment assignments. Together, this study provides the first global subcellular localization map of the Dictyostelium proteome, establishing a foundational resource for functional, cell biological, and evolutionary analyses. We are currently preparing manuscripts that further analyze (i) the membrane trafficking system of Dictyostelium , including detailed investigation of the contractile vacuole, Jotnarlogs, and patchy proteins, and (ii) the composition of the Dictyostelium peroxisome, with a focus on the subcellular localization of sterol biosynthesis enzymes. Investigators interested in using these data are encouraged to contact us prior to publication to avoid overlap and to facilitate coordinated and collaborative use of this resource.