Large pelagic sharks are declining in abundance in many oceans owing to fisheries exploitation. What is not known however is whether withinspecies geographical segregation of the sexes exacerbates this as a consequence of differential exploitation by spatially focused fisheries. Here we show striking sexual segregation in the fastest swimming shark, the shortfin mako Isurus oxyrinchus, across the South Pacific Ocean. The novel finding of a sexual 'line in the sea' spans a historical longline-fishing intensity gradient, suggesting that differential exploitation of the sexes is possible, a phenomenon which may underlie changes in the shark populations observed elsewhere.