title: The disulfide bond in chromogranin B, which is essential for its sorting to secretory granules, is not required for its aggregation in the trans-Golgi network.
abstract: Chromogranin B (secretogranin I), a protein sorted to secretory granules in many endocrine cells and neurons, undergoes selective aggregation during the sorting process in the trans-Golgi network. Reduction of the single, highly conserved intramolecular disulfide bond of chromogranin B by exposure of intact PC12 cells to the thiol reducing agent dithiothreitol has previously been shown to cause its missorting to the constitutive pathway of secretion. Using saponin perforation of membrane vesicles in aggregative buffer mimicking the milieu in the lumen of the trans-Golgi network (pH 6.4, 10 mM calcium), we show here that treatment with dithiothreitol does not prevent the aggregation of chromogranin B in this compartment. This implies that the loop in the chromogranin B polypeptide that is formed by the disulfide bond has a critical role in the membrane recognition of aggregated chromogranin B during secretory granule formation.
authors.affiliation: Institute for Neurobiology, University of Heidelberg, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Chanat
authors.forename: U
authors.initials: U
authors.lastname: Weiss
authors.forename: W B
authors.initials: WB
authors.lastname: Huttner
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002118:Calcium / Q000494:pharmacology; D002451:Cell Compartmentation / Q000187:drug effects; D002864:Chromogranins / Q000187:drug effects / Q000378:metabolism*; D003594:Cytoplasmic Granules / Q000378:metabolism*; D004220:Disulfides / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008954:Models, Biological; D010084:Oxidation-Reduction; D016716:PC12 Cells; D010446:Peptide Fragments / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship
chemical_list: D002864:Chromogranins; D004220:Disulfides; D010446:Peptide Fragments; D002118:Calcium; D004229:Dithiothreitol