Mis-sorting of procathepsin D in metastogenic tumor cells is not due to impaired synthesis of the phosphomannosyl signal

Synthesis, processing and sorting of pro-cathepsin D (proCD), the precursor of a lysosomal protease involved in tumor-cell proliferation and invasion, were compared in various human tumor cell lines. In cultures of HepG2, HT29 and MCF7 cells the extracellular CD activity was up to 10 times higher than in cultures of U937 and MG63 cells. Ammonium chloride, which disrupts the pH-dependent receptor-mediated transport of lysosomal enzymes, exerted a differential effect on the secretion of CD activity in these cells. As judged by metabolic labeling and immunoprecipitation, the secretion of CD synthesized in 24 hr in MG63 and U937 cells was enhanced 4- to 10-fold by ammonium chloride, while it was only slightly increased in HepG2, HT29 and MCF7 cells. In all tumor cells examined, a portion of proCD was segregated into the endosomal-lysosomal pathway in the presence of ammonium chloride. However, the intralysosomal maturation of CD was only inhibited by this drug in HT29 and MCF7 cells. The Golgi-associated processing of proCD, leading to the synthesis of uncovered phosphomannosyl groups, proceeded with comparable efficiency in all tumor cells examined. These results suggest that hypersecretion of proCD in HepG2, HT29 and MCF7 cells is linked to transport mechanisms, as yet unidentified, which are independent of mannose-6-phosphate.