Effects of chlorpromazine-pretreated melanin on lysosomal acid phosphatase and N-acetyl-beta-D-glucosaminidase in bovine ciliary body and iris in vitro.

We studied biochemically the effect of chlorpromazine-pretreated melanin on lysosomal enzyme activities in the bovine ciliary body and iris in vitro. Melanin was prepared from the bovine ciliary body and iris by acid treatment. Acid phosphatase and N-acetyl-beta-D-glucosaminidase of the ciliary body and iris were used as lysosomal marker enzymes. After the enzyme solution was incubated with melanin, enzyme activity was reduced and protein content in the supernatant was decreased. When melanin was pretreated with chlorpromazine, both enzyme activity and protein content in the supernatant remained higher than after the incubation with melanin alone, depending on the concentration of chlorpromazine. Chlorpromazine itself seemed to have little effect on the acid phosphatase and N-acetyl-beta-D-glucosaminidase at the concentration used in the incubation mixture. The increased enzyme activity, therefore, may result from a loss of the enzyme affinity for melanin after chlorpromazine pretreatment. These findings were discussed with respect to the binding mechanism of lysosomal enzymes to melanin and possible effect of chlorpromazine on biochemical interaction between lysosomal enzymes and melanin in vivo.