The demersal fish assemblages of the south-western Sea of Marmara and the north-eastern Aegean Sea were examined in relation to their biogeographical and environmental conditions. Sampling was carried out during bottom trawl surveys conducted during five periods between June 2006 and August 2007. A total of 40 hauls were processed, including 16 in the Sea of Marmara (38–74 m deep) and 24 in the north-eastern Aegean Sea (63–401 m deep). A total of 47,940 individuals, from 91 species and 47 families were collected. Cluster analysis of catch data indicated that there were three assemblages, one in the shelf of the Sea of Marmara and two in the Aegean Sea (<100 and >200 m depth). The numerical abundance and biomass of fish, mean fish weight, mean species richness, diversity and evenness were compared between these assemblages. Multivariate analysis was used to test differences in environmental conditions between areas, and relationships between species and environmental variables were explored by using redundancy analysis. In addition to bathymetric differences, differences in both diversity and species composition were observed between shelf assemblages in the two nearby areas. The distinct biogeographical, environmental characteristics (depth, dissolved oxygen and temperature) and fishing pressure are discussed as the main factors which could explain the differences detected.