Secretion and molecular forms of NESP55, a novel genomically imprinted neuroendocrine-specific protein from AtT-20 cells.
NESP55 (neuroendocrine secretory protein of M(r) 55,000) is a paternally imprinted proteoglycan, expressed specifically in endocrine cells and the nervous system. We investigated the subcellular localization and secretion of NESP55 in AtT-20 cells. NESP55 accumulated in the medium linearly over 24 h exceeding its intracellular content 3.7-fold by that time. Incubation of cells at 16 degrees C, to block protein export, inhibited basal secretion by 79%. Stimulation of AtT-20 cells with 8-Br-cAMP increased secretion of NESP55 by only 45%. The NESP55 secretory vesicles sedimented at a density of 1.2-1.4 M, which is slightly lighter than that of the large dense core vesicles. Immunofluorescence studies revealed immunoreactivity in the Golgi apparatus and a punctuate staining of processes or neurites. Our data demonstrate that NESP55 is mainly sorted to and released from a population of constitutive secretory vesicles, which are transported out of the perikarya into processes or axons. In addition, some NESP55 is also routed to the regulated pathway. The signal peptide of NESP55, as determined with peptide antisera, is 46 amino acids long and represents the best conserved region of this molecule suggesting that the signal peptide may have a function of its own. The subcellular localization and export of NESP55 from cells are reminiscent of neuronal proteoglycans forming the extracellular matrix, which are implicated in the development and maintenance of neuronal circuits and mechanisms of axonal guidance.
Eder
Susanne
S
Department of Pharmacology, Innsbruck Medical University, Innsbruck, Austria.
Leierer
Johannes
J
Klimaschewski
Lars
L
Wilhelm
Alexander
A
Volknandt
Walter
W
Laslop
Andrea
A
Fischer-Colbrie
Reiner
R
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002460:Cell Line; D002864:Chromogranins; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000235:genetics* / Q000378:metabolism*; D018392:Genomic Imprinting; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016716:PC12 Cells; D051381:Rats
D002864:Chromogranins; C406732:GNAS protein, human; C498644:GNAS1 protein, Bos taurus; C498642:Gnas protein, mouse; C498643:Gnas protein, rat; D019205:GTP-Binding Protein alpha Subunits, Gs