Latifah, 49, added a rap verse to the legendary musician's song, in which she added a line paying tribute to Bryant.
"Give love a shot / When you do, say 'Kobe' / 24 hours, eight days a week, trophies," she rapped.
The crowd could be heard cheering after the line.
The performance comes several weeks after Bryant died in a helicopter crash in California in late January. He was 41. Bryant's daughter Gianna was also among the victims of the crash.
On the day he passed, Latifah took to Instagram to share her grief.
The "Chicago" star shared a photo of a cross with the words "God Bless Kobe" written over it.
"Omg an icon and his daughter," she wrote in the caption. "Lord Find the Way to Make A Restful Peace For The Fans and The Family."
The NBA All-Star Game will take place Sunday evening and will feature a special tribute to Bryant by Jennifer Hudson before the game begins. Chaka Khan, Chance the Rapper, Lil Wayne, DJ Khaled and Quavo are also set to perform.