MATURATION OF HUMAN LACTASE-PHLORHIZIN HYDROLASE - PROTEOLYTIC CLEAVAGE OF PRECURSOR OCCURS AFTER PASSAGE THROUGH THE GOLGI-COMPLEX

Maturation of human intestinal lactase-phlorizin hydrolase (LPH) requires that a precursor (pro-LPH) be proteolytically processed to the mature microvillus membrane enzyme (m-LPH). The subcellular site of this processing is unknown. Using low-temperature experiments and brefeldin A (BFA), intracellular transport was blocked in intestinal epithelial cells. In Caco-2 cells incubated at 18-degrees-C, pro-LPH was complex-glycosylated but not cleaved, while at 20-degrees-C small amounts of proteolytically processed LPH were observed. These data exclude a pre-Golgi proteolytic event. BFA completely blocked proteolytic maturation of LPH and lead to an aberrant form of pro-LPH in both Caco-2 cells and intestinal "plants. Therefore, proteolytic processing of LPH is a post-Golgi event, occuring either in the trans-Golgi network, transport vesicles, or after insertion of pro-LPH into the microvillus membrane.