Control of pancreatic bile-salt-dependent-lipase secretion by the glucose-regulated protein of 94 kDa (Grp94).
Bile-salt-dependent lipase (BSDL; EC 3.1.1.13) is an enzyme expressed by the pancreatic acinar cell and secreted as a component of the pancreatic juice. During its route towards secretion, BSDL is associated with intracellular membranes by means of a multiprotein folding complex, which includes the glucose-regulated protein of 94 kDa (Grp94). We have postulated that the association of BSDL with membranes is required for the complete O-glycosylation of the protein, which diverts BSDL from a degradation route and consequently allows its secretion. To further characterize the role of Grp94 in BSDL secretion, we have studied the effect of a ribozyme specifically targeted to Grp94 mRNA. This ribozyme has been transfected into AR4-2J cells, and we have shown that a decrease in Grp94 expression leads to a concomitant decrease in BSDL secretion and expression. Geldanamycin (GA), which alters Grp94 functions, also affects the release of BSDL into the culture medium of AR4-2J cells. BSDL expressed in GA-treated AR4-2J cells is unstable. Furthermore, under conditions that decrease the level of BSDL secretion, no intracellular accumulation of the enzyme was observed, suggesting that BSDL that cannot associate with (or be structured by) Grp94 could be rapidly degraded. We have further shown that this degradation probably occurs via the ubiquitin-dependent pathway. Altogether, these results indicate that Grp94 has a pivotal role in BSDL folding and in the sorting of this pancreatic enzyme.
Nganga
A
A
INSERM Unité 260-Faculté de Médecine-Timone, 27 bld Jean Moulin, 13385 Marseille, Cedex 05, France.
Bruneau
N
N
Sbarra
V
V
Lombardo
D
D
Le Petit-Thevenin
J
J
D000818:Animals; D001483:Base Sequence; D016227:Benzoquinones; D002087:Butyrates / Q000494:pharmacology; D015536:Down-Regulation; D004795:Enzyme Stability / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D047029:Lactams, Macrocyclic; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D010179:Pancreas / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D011809:Quinones / Q000494:pharmacology; D016337:RNA, Catalytic / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D002787:Sterol Esterase / Q000378:metabolism*; D013379:Substrate Specificity; D014162:Transfection; D014407:Tumor Cells, Cultured; D014452:Ubiquitins / Q000378:metabolism
D016227:Benzoquinones; D002087:Butyrates; D018840:HSP70 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D008565:Membrane Proteins; D011809:Quinones; D016337:RNA, Catalytic; D012333:RNA, Messenger; D014452:Ubiquitins; C018551:glucose-regulated proteins; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase; C001277:geldanamycin