Intracellular transport of secretory proteins in the pancreatic exocrine cell. IV. Metabolic requirements.
Since in the pancreatic exocrine cell synthesis and intracellular transport of secretory proteins can be uncoupled (1), it is possible to examine separately the metabolic requirements of the latter process. To this intent, guinea pig pancreatic slices were pulse labeled with leucine-(3)H for 3 min and incubated post-pulse for 37 min in chase medium containing 5 x 10(-4)M cycloheximide and inhibitors of glycolysis, respiration, or oxidative phosphorylation. In each case, the effect on transport was assessed by measuring the amount of labeled secretory proteins found in zymogen granule fractions isolated from the corresponding slices. This assay is actually a measure of the efficiency of transport of secretory proteins from the cisternae of the rough endoplasmic reticulum (RER) to the condensing vacuoles of the Golgi complex which are recovered in the zymogen granule fraction (16). The results indicate that transport is insensitive to glycolytic inhibitors (fluoride, iodoacetate) but is blocked by respiratory inhibitors (N(2), cyanide, Antimycin A) and by inhibitors of oxidative phosphorylation (dinitrophenol, oligomycin). Except for Antimycin A, the effect is reversible. Parallel radioautographic studies and cell fractionation procedures applied to microsomal subfractions have indicated that the energy-dependent step is located between the transitional elements of the RER and the small, smooth-surfaced vesicles at the periphery of the Golgi complex. Radiorespirometric data indicate that the substrates oxidized to support transport are endogenous long-chain fatty acids.
D000818:Animals; D000963:Antimetabolites / Q000494:pharmacology*; D001345:Autoradiography; D001693:Biological Transport, Active / Q000187:drug effects*; D003513:Cycloheximide / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000502:physiology; D006056:Golgi Apparatus / Q000502:physiology; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007930:Leucine / Q000378:metabolism; D008854:Microscopy, Electron; D010101:Oxygen Consumption / Q000187:drug effects; D010179:Pancreas / Q000166:cytology* / Q000378:metabolism; D014176:Protein Biosynthesis; D013696:Temperature; D013997:Time Factors
D000963:Antimetabolites; D003513:Cycloheximide; D007930:Leucine