Cell type-specific storage of dopamine beta-monooxygenase.
Expression of dopamine beta-monooxygenase (DBM), the enzyme that converts dopamine into norepinephrine, is limited to adrenal chromaffin cells and a small population of neurons. We studied DBM trafficking to regulated granules by stably expressing rat DBM in AtT-20 corticotrope tumor cells, which contain regulated granules, and in Chinese hamster ovary (CHO) cells, which lack regulated granules. The behavior of exogenous DBM in both cell lines was compared with endogenous DBM in adrenal chromaffin cells. CHO cells secreted active DBM, indicating that production of active enzyme does not require features unique to neuroendocrine cells. Pulse-chase experiments indicated that early steps in DBM maturation followed a similar time course in AtT-20, CHO, and adrenal chromaffin cells. Use of a conformation-sensitive DBM antiserum indicated that acquisition of a folded structure occurred with a similar time course in all three cell types. Cell type-specific differences in DBM trafficking became apparent only when storage in granules was examined. As expected, DBM was stored in secretory granules in chromaffin cells; CHO cells failed to store DBM. Despite the fact that AtT-20 cells have regulated granules, exogenous DBM was not stored in these granules. Thus storage of DBM in secretory granules requires cell type specific factors.
Oyarce
A M
AM
Department of Neuroscience, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2105, USA.
Eipper
B A
BA
D000818:Animals; D001692:Biological Transport; D016466:CHO Cells; D006224:Cricetinae; D003594:Cytoplasmic Granules / Q000378:metabolism; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism*; D009928:Organ Specificity; D051381:Rats
D004299:Dopamine beta-Hydroxylase