The major surfactant protein SP-A is thought to play an important role in the regulation of the structure and the metabolism of pulmonary surfactant. We investigated whether this protein may be a target in ozone toxicity. Several properties were studied that are probably relevant to the physiological functions of SP-A. In vitro exposure of human and canine SP-A to ozone led to decreases in 1) self-association of SP-A, 2) SP-A-mediated lipid aggregation, and 3) binding of SP-A to immobilized mannose. SP-A was neither degraded nor cross-linked by ozone exposure. Exposure of canine SP-A to ozone led to an increase in the apparent molecular weight of monomeric SP-A. Human SP-A did not show this change. Oxidation of methionine and tryptophan residues in canine SP-A was detected following ozone exposure. Reactions of ozone with other amino acid residues were not observed. The impairments of structure and properties of SP-A may contribute to the toxic action of ozone in the lungs.