Phosphorylation of the rat pancreatic bile-salt-dependent lipase by casein kinase II is essential for secretion.
Bile-salt-dependent lipase (BSDL, EC 3.1.1.-) is an enzyme expressed by the pancreatic acinar cells and secreted as a component of the pancreatic juice of all examined species. During its secretion route BSDL is associated with intracellular membranes. This association allows the complete glycosylation of the enzyme or participates in the inhibition of the enzyme activity, which can deleterious for the acinar pancreatic cell. Thereafter, the human BSDL is phosphorylated by a serine/threonine protein kinase and released from intracellular membranes. In the present study, we show that the rat pancreatic BSDL, expressed by AR4-2J cells used as a model, is phosphorylated by a protein kinase that is insensitive to inhibitors of protein kinases A, C or G and that the phosphorylation process is favoured by okadaic acid (an inhibitor of protein phosphatases 1 and 2A). However, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole (DRB), which is a specific inhibitor of casein kinase II, abolishes the phosphorylation in vitro of BSDL within micro- somes of AR4-2J pancreatic cells. We showed further that the alpha-subunit of casein kinase II co-locates with BSDL within the lumenal compartment of the Golgi. Genistein, which perturbs the trans-Golgi network, also inhibits the phosphorylation of BSDL, suggesting that this post-translational modification of BSDL probably occurred within this cell compartment. The inhibition of the phosphorylation of BSDL by DRB also decreases the rate at which the enzyme is secreted. Under the same conditions, the rate of alpha-amylase secretion was not modified. These data strongly suggest that phosphorylation is a post-translational event, which appears to be essential for the secretion of BSDL.
Pasqualini
E
E
INSERM U.260, Faculté de Médecine/Timone, 27 bld Jean Moulin, 13385 Marseille cedex 05, FRANCE.
Caillol
N
N
Valette
A
A
Lloubes
R
R
Verine
A
A
Lombardo
D
D
D000818:Animals; D047390:Casein Kinase II; D002451:Cell Compartmentation; D002460:Cell Line; D004004:Dichlororibofuranosylbenzimidazole / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D010179:Pancreas / Q000201:enzymology*; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D051381:Rats; D002787:Sterol Esterase / Q000737:chemistry / Q000378:metabolism*; D000516:alpha-Amylases / Q000378:metabolism
D004791:Enzyme Inhibitors; D004004:Dichlororibofuranosylbenzimidazole; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase; D000516:alpha-Amylases