title: The apical sorting of lactase-phlorizin hydrolase implicates sorting sequences found in the mature domain.
abstract: Polarized transport of proteins is contingent on the presence of specific protein structures or motifs that function as sorting signals. Our model protein to analyze and to identify such signals is that of lactase-phlorizin hydrolase (LPH), a strictly polarized brush border membrane protein of small intestinal epithelial cells. It is synthesized as a large pro-LPH precursor molecule, which is proteolytically processed to yield the mature brush border enzyme (LPHbeta). Pro-LPH as well as LPHbeta are correctly sorted to the brush border membrane. In this paper we examine the location of putative sorting signals in the pro-LPH molecule. Expression of a cDNA encoding the LPHbeta mature form in the absence of the LPHalpha species in Madin-Darby canine kidney (MDCK) cells reveal an LPHbeta molecule that is not as transport-competent as wild type pro-LPH. The proportion of complex glycosylated LPHbeta constitutes not more than 10% of the total synthesized protein. This form displays a similar trypsin sensitive pattern as wild type intestinal LPHbeta suggesting comparable folding patterns of the two species. Complex glycosylated LPHbeta is sorted to the apical membrane more efficiently than wild type pro-LPH. We conclude that the apical sorting signals for pro-LPH are exclusively found in the LPHbeta mature domain.
authors.affiliation: Institute of Microbiology, Heinrich Heine University of Düsseldorf, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Jacob
authors.forename: K P
authors.initials: KP
authors.lastname: Zimmer
authors.forename: H
authors.initials: H
authors.lastname: Naim
authors.forename: H Y
authors.initials: HY
authors.lastname: Naim
mesh_terms: D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D004285:Dogs; D015870:Gene Expression; D006031:Glycosylation; D006801:Humans; D010694:Lactase-Phlorizin Hydrolase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D010694:Lactase-Phlorizin Hydrolase