title: Localization of integral membrane peptidylglycine alpha-amidating monooxygenase in neuroendocrine cells.
abstract: Peptidylglycine alpha-amidating monooxygenase (PAM) catalyzes the amidation of glycine-extended peptides in neuroendocrine cells. At steady state, membrane PAM is accumulated in a perinuclear compartment. We examined the distribution of membrane PAM in stably transfected AtT-20 cells and compared its localization to markers for the trans-Golgi network (TGN), endosomes, and lysosomes. At the light microscopic level, the distribution of membrane PAM does not overlap extensively with lysosomal markers but does overlap with TGN38 and with SCAMP, a component of post-Golgi membranes involved in recycling pathways. By immunoelectron microscopy, membrane PAM is present in tubulovesicular structures which constitute the TGN; some of these PAM-containing tubulovesicular structures are more distal to the Golgi stacks and do not contain TGN38. While some POMC-derived peptides are present in tubulovesicular structures like those that contain membrane PAM, the majority of the POMC-derived peptides are present in secretory granules. There is little overlap between the steady state distribution of membrane PAM and internalized FITC-transferrin in the early endosomes. Few of the perinuclear PAM-containing structures are labeled with HRP or WGA-HRP even following long incubations. Therefore, membrane PAM is localized to perinuclear tubulovesicular structures which are partially devoid of TGN38 and are not all endosomal in origin.
authors.affiliation: Physiology Department, The University of North Carolina at Chapel Hill 27599, USA.
authors.forename: S L
authors.initials: SL
authors.lastname: Milgram
authors.forename: S T
authors.initials: ST
authors.lastname: Kho
authors.forename: G V
authors.initials: GV
authors.lastname: Martin
authors.forename: R E
authors.initials: RE
authors.lastname: Mains
authors.forename: B A
authors.initials: BA
authors.lastname: Eipper
mesh_terms: D000302:Adrenal Cortex / Q000166:cytology / Q000201:enzymology*; D002451:Cell Compartmentation; D002460:Cell Line; D004705:Endocytosis; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009097:Multienzyme Complexes
chemical_list: D009097:Multienzyme Complexes; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase