title: Neutrophil elastase sorting involves plasma membrane trafficking requiring the C-terminal propeptide.
abstract: The primary granules/secretory lysosomes of neutrophils store mature neutrophil elastase (NE) as a luminal protein after proteolytic removal of N-terminal and C-terminal pro-peptides from a proform of NE. The N-terminal pro-peptide prevents premature activation that might be toxic to the cell, but the C-terminal pro-peptide has no defined function. In this study, we investigated the role of the C-terminal pro-peptide in trafficking of NE by expressing, in rat basophilic leukemia (RBL) cells, both wild-type NE and the mutant NE/Delta248-267, which lacks the C-terminal pro-peptide. Both transfected proteins were found to be targeted to secretory lysosomes. In addition, results from antibody ligation and cell-surface biotinylation indicated that proform of NE was targeted to the plasma membrane, and then subjected to endocytosis. The results were supported by the detection of targeting of the proform to the plasma membrane followed by internalization both in RBL cells and normal granulopoietic precursor cells. Targeting of NE to the plasma membrane required the C-terminal pro-peptide as NE/Delta248-267 expressed in RBL cells bypassed plasma membrane trafficking. Our results indicate targeting of a population of NE to the plasma membrane and internalization dependent on the C-terminal NE pro-peptide.
authors.affiliation: Department of Clinical Sciences, Section for Clinical and Experimental Infection Medicine, B14, BMC, SE-221 84 Lund, Sweden.
authors.forename: Hans
authors.initials: H
authors.lastname: Tapper
authors.forename: Linda
authors.initials: L
authors.lastname: Källquist
authors.forename: Ellinor
authors.initials: E
authors.lastname: Johnsson
authors.forename: Ann-Maj
authors.initials: AM
authors.lastname: Persson
authors.forename: Markus
authors.initials: M
authors.lastname: Hansson
authors.forename: Inge
authors.initials: I
authors.lastname: Olsson
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D019272:Leukocyte Elastase / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D051381:Rats
chemical_list: D015415:Biomarkers; D010455:Peptides; D011498:Protein Precursors; D019272:Leukocyte Elastase