PERTURBATION OF REGULATED SECRETION IN THE PANCREATIC ACINAR CELL-LINE, AR42J

被引:18
作者
SACHS, E [1 ]
JAMIESON, JD [1 ]
机构
[1] YALE UNIV,SCH MED,DEPT CELL BIOL,333 CEDAR ST,NEW HAVEN,CT 06510
来源
AMERICAN JOURNAL OF PHYSIOLOGY | 1992年 / 262卷 / 02期
关键词
INTRACELLULAR PROTEIN TRANSPORT; SECRETORY GRANULES; CHOLECYSTOKININ; EXOCYTOSIS; CONSTITUTIVE SECRETION; AR42J CELLS;
D O I
10.1152/ajpgi.1992.262.2.G257
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
The regulated secretory pathway comprises accelerated discharge of proteins in response to hormonal stimuli, their presence in secretory granules (SG), and a long intracellular residence time. Dexamethasone induction of AR42J results in an increase in granule content and responsiveness to cholecystokinin (CCK). We studied the effects of conditions implicated in sorting of secretory proteins into the regulated pathway using [S-35]methionine pulse-chase protocols that examine transport of secretory proteins from the rough endoplasmic reticulum (RER) --> SG and specifically from the Golgi complex (GC) --> SG. The latter uses a chase at 20-degrees-C to allow accumualation of labeled proteins in the trans-Golgi, followed by a shift to 37-degrees-C that initiates their transport to SG under test conditions. Quantitation of CCK-8-stimulated discharge of prestored amylase and of newly synthesized labeled proteins that have entered SG during the chase enables us to examine the effect of perturbants over selected parts of the pathway. The effects of acidic intracellular compartments, the cytoskeleton, protein synthesis, ATP, and temperature on pre- and post-Golgi entry of proteins into the regulated pathway were studied. NH4Cl, monensin, Na azide, incubation at 20-degrees-C, and pertussis toxin retarded RER --> SG transport without affecting amylase discharge. Only incubation with 20 mM NH4Cl or 1-mu-M monensin inhibited transfer of newly synthesized proteins from the late GC --> SG. RER --> Golgi or intra-Golgi transport thus appears to require ATP and possibly guanosine 5'-triphosphate (GTP)-binding proteins. Acidic compartments appear to be essential for sorting of secretory proteins from the GC --> SG.
引用
收藏
页码:G257 / G266
页数:10
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