Effect of feeding riboflavin deficient diet to rats on lens protein composition was investigated. Total proteins and profile of soluble and insoluble proteins in lenses from rats fed on a riboflavin deficient diet for seven weeks were found to be similar to that of the vitamin supplemented diet. Distribution of high molecular weight protein (above 4 X 10(6) daltons) isolated from the 9,900 g supernatant fraction was found to be significantly higher and gamma crystallin was significantly lower in riboflavin deficient group as compared to the normal lenses. However, the distribution of alpha and beta crystallins was not affected. Gel electrophoresis in sodium dodecyl sulfate of soluble lens proteins from riboflavin deficient animals had lower proportions of polypeptide species with molecular weight above 40,000 daltons while insoluble protein fraction had higher proportions of these polypeptide species as compared to control rats. These data suggest that the composition of lens proteins is altered in riboflavin deficiency.