The kinetics of the reactions between fluorescently labeled sperm whale myoglobin-(110-121) peptide and the murine major histocompatibility complex class II protein I-Ed have been analyzed. The presence in solution of both short- and long-lived protein-peptide complexes is demonstrated by the biphasic dissociation of the myoglobin peptide from I-Ed. The formation of the long-lived terminal complex is preceded by a characteristic induction phase. It is shown that the initially formed complex of the myoglobin peptide and I-Ed is a kinetic intermediate that undergoes a unimolecular reaction to form the terminal complex. Reactions between peptides and the class II proteins thus involve an intermediate structurally distinct from the terminal complex. The terminal complex presumably has a structure that is biologically active and similar to the published class II protein-peptide crystal structure.