ABSTRACT
The complex mechanism of intracellular transport is regulated by free calcium in different manners. Calcium binding proteins regulate several aspects of the vesicle fusion mechanism mediated by NSF (N-ethylmaleimide sensitive fusion factor). At least in some regulated exocytosis, calcium-binding proteins are the trigger for fusion downstream of NSF, Still, calcium-binding proteins, such as annexins, may be part of a different fusion mechanism mediating some specific transport steps or working in parallel to the NSF-dependent fusion process. Calcium is not the only ion necessary for the function of factors involved in vesicular transport. A zinc requirement has been also proposed. One of the zinc-dependent factors is probably a protein with a cysteine-rich region that coordinates zinc and binds phorbol esters. Although protein kinase C is the more prominent family of proteins carrying this domain, the factor necessary for transport does not appear to function as a kinase.
Subject(s)
Animals , Dogs , Biological Transport , Calcium , Calcium-Binding Proteins , Metalloproteins/physiology , Zinc , Cell Line , Phorbol Esters/metabolism , Exocytosis , Kidney , Intracellular Fluid/metabolism , Membrane Fusion , Protein Binding , Protein Kinase C/physiology , Carrier Proteins/physiology , Coated Vesicles/physiologyABSTRACT
Adrenocorticotropic hormone (ACTH) triggers well-defined responses in Y-1 cells. Among them is steroidogenesis stimulation. We have previously shown that phorbol 12-myristate 13-acetate (PMA), an activator of the calcium- and phospholipid-dependent protein kinase (PKC) is able to mimic all the responses triggered by ACTH in these cells, including steroidogenesis stimulation. Short (2 h) treatment with PMA leads to only 20-30 per cent of the maximal steroidogenesis stimulation obtained with ACTH. However, the steroid secretion in the 2 h that follows the short-term (2 h) PMA treatment reaches the same levels as observed with ACTH, i.e., a 12- to 15-fold increase. We also show that this effect is restricted to cells treated with PMA for up to 4 h, while treatment for longer periods of time causes a reduction of the steroid biosynthesis rate, an effect that is not observed in cells treated with ACTH or N6,2'-O-dibutyryladenosine 3',5'-cyclic monophosphate (dcAMP). These results suggest that activation of PKC can elicit the first phase of ACTH steroidogenesis stimulation, but not the second one, which strictly depends on activation of cAMP-dependent protein kinase.
Subject(s)
Adrenocorticotropic Hormone/metabolism , Cyclic AMP/metabolism , Phorbol Esters/metabolism , Steroids/biosynthesis , Adrenocorticotropic Hormone/physiologyABSTRACT
La activación de alfa2-adrenoreceptores o Ri-adenosinreceptores en células adiposas disminuyó considerablemente la acumulación de AMP cíclico inducido por el agonista beta-adrenérgico isoprenalina. Esta acción casi fue abolida en células de hamster tratadas con toxina de la tosferina. El ester forbol activo, forbol 12-miristrato 13 acetato, fue incapaz de reproducir el efecto de la toxina en este modelo. Los datos indican que la activación de la proteinoquinasa C en este modelo celular no altera la rama inhibitoria de la adenil ciclasa