The crash happened at 6:12 pm in the area of Cesar Chavez Boulevard and 3rd Street.
Fresno police say the woman stepped into the roadway from the center median and was hit by an oncoming vehicle.
She was taken to a local hospital, where she later died.
The driver pulled over after the crash and is cooperating with officers.
The eastbound lanes of Cesar Chavez Boulevard have been closed between 2nd Street and 3rd Street as the investigation continues.