The protagonist of Ma Rainey’s Black Bottom is the titular character, a blues singer whose recording session is the setting of August Wilson’s play. However, the other musicians who take part in its debates about race and economics play significant parts in the narrative.

In some respects, the antagonist is Levee, the youngest of the musicians, who wants to get together his own band to record his own songs, and whose faith that the white managers who control the music business will allow him to succeed is dashed. His frustration leads him to stab another of the musicians, an event that represents the violence black people do to each other because they cannot reach the white oppressors who are the real targets of their rage.

However, the real antagonist is the white power structure that controls the music business and the country. The play’s dialogue is largely about the characters’ experiences of racism and their knowledge that white managers are using their talent to make large amounts of money while paying them little.

That power structure is represented by the three white characters:

Sturdyvant, the owner of the recording studio, cares only about money. He is uncomfortable with black people and treats them with thinly veiled contempt. He precipitates the violent confrontation by offering Levee an insultingly small sum for his songs.

Irvin is Ma Rainey’s opportunistic manager, who prides himself on being able to interface with blacks and essentially makes a living by acting as a go-between for black talent and white owners of music businesses.

A policeman enters with Ma Rainey and two of her sidekicks, arguing about an automobile accident Ma may have been responsible for. Once satisfied she is “important,” he takes a bribe from Irvin and leaves.

None of these characters operates precisely as an antagonist, although Sturdyvant comes closest to playing that role.

The protagonist in Ma Rainey's Black Bottom is Ma Rainey herself. The musicians wait around until Ma Rainey, an African-American singer who is regarded as the "mother of the blues," arrives at their studio in the South Side of Chicago in the 1920s. The musicians--Levee, Toledo, Slow Drag, and Cutler-- wait and discuss their frustrations dealing with a society that is filled with racial prejudice.

The antagonist is Levee, a trumpet player, who places hope in the idea that his music will help him gain respect from the white world that controls the music industry. Ma Rainey, for her part, knows that her power only comes from her ability to make money for her white managers. As a result, she is demanding, requesting respect until her days making music are over. The dramatic conflict is that Levee wants the producers to make his record, but he is crushed when the white producer, Sturdyvant, refuses and when Ma Rainey fires him. When Toledo mistakenly steps on Levee's shoes, Levee stabs him. Toledo is a supporter of the self-determination of African-Americans, so his stabbing is symbolic of Levee's rejection of this idea in favor of winning (and failing to win) respect from white people.