Launch from Cape Canaveral (KSC); landing on Edwards AFB.

STS-53 was the final dedicated Department of Defense mission. Discovery carried a classified primary payload for the United States Department of Defense, two unclassified secondary payloads and nine unclassified middeck experiments. Discovery's primary payload, USA-89 NSSDC ID 1992-086B is also known as "DoD-1", and was the shuttle's last major payload for the Department of Defense. The satellite was the second launch of a Satellite Data System-2 military communications satellite, after USA-40 on STS-28.

Secondary payloads contained in or attached to Get Away Special (GAS) hardware in the cargo bay included the Orbital Debris Radar Calibration Spheres (ODERACS) the combined Shuttle Glow Experiment/Cryogenic Heat Pipe Experiment (GCP). Middeck experiments included Microcapsules in Space (MIS-l); Space Tissue Loss (STL); Visual Function Tester (VFT-2); Cosmic Radiation Effects and Activation Monitor (CREAM); Radiation Monitoring Equipment (RME-III); Fluid Acquisition and Resupply Experiment (FARE); Hand-held, Earth-oriented, Real-time, Cooperative, User-friendly, Location-targeting and Environmental System (HERCULES); Battlefield Laser Acquisition Sensor Test (BLAST); and the Cloud Logic to Optimize Use of Defense Systems (CLOUDS).

The experiments ODERACS, BLAST and HERCULES were reduced and delayed respectively because of battery problems.