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Exit of nonglycosylated secretory proteins from the rough endoplasmic reticulum is asynchronous in the
exocrine pancreas
The path and synchrony of intracellular transport of 12 secretory proteins of the guinea pig exocrine
pancreas have been studied in pulse-chase amino acid labeling experiments by quantitative analysis of the
individual proteins recovered in subcellular fractions and extracellular samples. Protein fractionation was
accomplished by two-dimensional isoelectric focusing/SDS-gel electrophoresis. Use of a double-label protocol
allowed correction of the data on a protein-by-protein basis for leakage and adsorption artifacts which
accompany tissue homogenization. All the labeled secretory (pro)enzymes, including their isoenzymic forms,
were recovered in rough microsomal, Golgi-enriched and granule fractions during their transport to the cell
surface. However, major asynchrony was observed at four levels: exit from the rough endoplasmic reticulum;
transit through the Golgi complex; entry into granules; and discharge from the cell. Rapid transport rates were
observed for trypsinogen, chymotrypsinogen 2, procarboxypeptidase A2, and lipase 2. Slow transport rates were
observed for amylase and procarboxypeptidase B. In the presence of carbamylcholine or cholecystokinin
stimulation, the times required for 40% discharge of labeled chymotrypsinogen 2, trypsinogen, amylase, and
procarboxypeptidase B were 98, 102, 148, and 180 min, respectively. Transport rates did not correlate with
isoelectric point, molecular weight, or the presence of carbohydrate. These data suggest that interactions occur
within the rough endoplasmic reticulum, either between secretory (nonglyco)-proteins themselves or between
such proteins and the cisternal face of the rough endoplasmic reticulum.
Rough ER and Pancreatic Cells: Condensation-sorting events in the rough endoplasmic reticulum of
exocrine pancreatic cells.
In guinea pig exocrine pancreatic cells intracisternal granules (ICGs) occur at a low frequency within the
lumen of the RER. By starving and refeeding guinea pigs or injecting them in CoCl2 solution, the number of
these granules is greatly increased. We show here that ICGs contain the complete set of secreted pancreatic
digestive enzymes and proenzymes. Two other soluble proteins in the lumen of the RER, GRP 78/BiP and
protein disulphide isomerase (PDI), are specifically excluded from ICGs. The formation of ICGs, which occurs
without acidification of the RER cisternae, is therefore a sorting event involving the co-condensation of a
complete set of secretory enzymes and proenzymes, which for brevity we refer to collectively as the zymogens.
With the exception of approximately 50% of the RNase, the zymogens in ICGs are covalently cross-linked by
intermolecular disulphide bonds. The synthesis of all three resident ER cisternal proteins--PDI, GRP 78/BiP,
and GRP 94--with the carboxy-terminal sequence KDEL, is induced in response to the accumulation of massive
amounts of misfolded secretory protein in the ICGs in the lumen of the RER. After injection of rats with large
doses of parachlorophenylalanine-methylester, crystals form in the lumen of the RER. We show that these
crystals appear to be a lattice of amylase with the other zymogens incorporated between the layers. Both GRP
78/BiP and PDI are excluded from these crystals. The formation of these amylase crystals within the RER and
the inclusion of other zymogens is, therefore, also a sorting event. These data establish that in exocrine
pancreatic cells zymogens can co-condense in the RER into either amorphous aggregates or crystals that
exclude other soluble RER proteins. This demonstrates that co-condensation is a mechanism capable of sorting
zymogens within the secretory pathway.
Golgi Apparatus and Epithelial Cells: THE FUNCTION OF THE GOLGI COMPLEX IN
TRANSITIONAL EPITHELIUM
The superficial squamous cells of rat transitional epithelium are limited, on their luminal face, by an
asymmetrically thickened membrane. Patches of similar thick membrane are found in the walls of the Golgi
cisternae and it is suggested that the Golgi system is the site of assembly of the thick plasma membrane. This
implies membrane flow from the Golgi apparatus to the cell surface, and there is indirect evidence that the
membrane is transported in the form of fusiform vacuoles, derived from the Golgi cisternae, which fuse with,
and become part of, the free cell membrane. Uptake of injected Imferon shows that similar, large, thick-walled
vacuoles may be formed by invagination of the free cell surface. Some of these vacuoles are subsequently
transformed into multivesicular bodies and autophagic vacuoles. The formation of other large heterogeneous
bodies is described, and some of these are shown to have acid phosphatase activity.