Membrane dipeptidase and glutathione are major components of pig pancreatic zymogen granules.
Membrane proteins of highly purified porcine zymogen granules were separated by two-dimensional gel electrophoresis in order to isolate proteins which are involved in intracellular trafficking of digestive enzymes in the exocrine pancreas. A 48-kDa glycoprotein was a major component in membrane preparations washed with 0.1 M Na2CO3 and 0.5 M NaCl. By N-terminal amino acid sequencing this protein was identified as membrane dipeptidase (MDP; EC 3.4.13.19). MDP mRNA levels in rat pancreas were increased threefold by feeding rats with FOY-305, which is a known stimulus of endogenous cholecystokinin release from the gut. Cholecystokinin then stimulates secretion in pancreatic acinar cells. In another set of experiments treatment of the rat pancreatic acinar tumor cell line AR42J with dexamethasone led to an eightfold increase in the expression of MDP. Thus, the expression pattern of the MDP gene in response to hormonal stimulation in vivo and in vitro resembles those found for most of the enzymes and proteins which are involved in secretion. Since MDP has been thought to have a role in glutathione (GSH) metabolism, we also measured GSH concentration in zymogen granules and found high levels of GSH. Based on our data we propose a working model for the function of MDP. According to this model, MDP might play a pivotal role in maintaining the oxidizing conditions in the ER, which are required for the correct folding of secretory proteins.
Höfken
T
T
Department of Gastroenterology, Philipps University, Marburg, 35032, Germany.
Linder
D
D
Kleene
R
R
Göke
B
B
Wagner
A C
AC
D000595:Amino Acid Sequence; D000818:Animals; D003594:Cytoplasmic Granules / Q000201:enzymology*; D004150:Dipeptidases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D004792:Enzyme Precursors / Q000737:chemistry*; D005786:Gene Expression Regulation; D005978:Glutathione / Q000737:chemistry* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000201:enzymology; D008297:Male; D008969:Molecular Sequence Data; D010179:Pancreas / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D017208:Rats, Wistar; D013552:Swine; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; D004150:Dipeptidases; C038937:dipeptidase; D005978:Glutathione