Energy utilization by the flagellum of motile sea urchin sperm is tightly coupled to the rate of energy production by the mitochondrion. This tight coupling depends upon the transport of high energy phosphate (P) from mitochondrion to axoneme, which we propose to be mediated by a phosphorylcreatine shuttle. The shuttle employs distinct mitochondrial and axonemal creatine kinase isozymes, the latter being a novel creatine kinase of 145 kd. To examine whether P is directed to the tail by such a shuttle, we inactivated creatine kinase specifically with fluorodinitrobenzene. Creatine kinase inactivation led to an inhibition of coupled, but not uncoupled, respiration and affected the pattern of sperm motility as predicted for the disruption of an obligatory link in P transport.