This project is designed to study several aspects of lysosomal hydrolase (beta-hexosaminidase) transport in normal and I-cell disease fibroblasts. These studies include the quantitative (intra- and extracellular distribution) and qualitative (electrophoretic mobility, lectin binding affinity, fibroblast uptake properties) changes in beta-hexosaminidase induced by known excretion effectors. These effectors include the Na ion/K ion ionophores monensin, ouabain and nigericin; the CA ions ionophore, A23187; the oxidative phosphorylation inhibitor, carbonyl cyanide m-chlorophenylhydrazone (CCCP); and the amine chloroquine. Studies will also include subcellular fractionation and localization experiments in fibroblasts for the purpose of locating individual isoenzymes in specialized vesicles and for tracking the migration of these isoenzymes from the site of synthesis to lysosomes or to excretion.