ATP-consuming and ATP-generating enzymes secreted by pancreas.
Pancreatic acini release ATP in response to various stimuli, including cholecystokinin octapeptide (CCK-8), as we show in the present study. There were indications that pancreatic juice also contains enzymes that could hydrolyze ATP during its passage through the ductal system. The aim of this study was to determine which ATP-degrading and possibly ATP-generating enzymes were present in pancreatic secretion. For this purpose, pancreatic juice was collected from anesthetized rats stimulated with infusion of CCK-8. Purine-converting activities in juice samples were assayed by TLC using either [gamma-(32)P]ATP or (14)C/(3)H-labeled and unlabeled nucleotides as appropriate substrates. Data show that the juice contains the enzyme ecto-nucleoside triphosphate diphosphohydrolase that can hydrolyze both [(14)C]ATP and [(3)H]ADP about equally well, i.e. CD39. Reverse-phase high-performance liquid chromatography analysis additionally shows that this enzyme has broad substrate specificity toward other nucleotides, UTP, UDP, ITP, and IDP. In addition, secretion contains ecto-5'-nucleotidase, CD73, further converting [(3)H]AMP to adenosine. Along with highly active hydrolytic enzymes, there were also ATP-generating enzymes in pancreatic juice, adenylate kinase, and NDP kinase, capable of sequentially phosphorylating AMP via ADP to ATP. Activities of nonspecific phosphatases, nucleotide pyrophosphatase/phosphodiesterases, and adenosine deaminase were negligible. Taken together, CCK-8 stimulation of pancreas causes release of both ATP-consuming and ATP-generating enzymes into pancreatic juice. This newly discovered richness of secreted enzymes underscores the importance of purine signaling between acini and pancreatic ducts lumen and implies regulation of the purine-converting enzymes release.
Yegutkin
Gennady G
GG
MediCity Research Laboratory and Department of Medical Microbiology, Turku University and National Public Health Institute, FIN-2050 Turku, Finland.
Samburski
Sergei S
SS
Jalkanen
Sirpa
S
Novak
Ivana
I
D015720:5'-Nucleotidase / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000096:biosynthesis* / Q000378:metabolism* / Q000502:physiology; D000263:Adenylate Kinase / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD; D001081:Apyrase / Q000378:metabolism; D002478:Cells, Cultured; D005260:Female; D009701:Nucleoside-Diphosphate Kinase / Q000378:metabolism; D046790:Pancreas, Exocrine / Q000201:enzymology* / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D015703:Antigens, CD; D000255:Adenosine Triphosphate; D000263:Adenylate Kinase; D009701:Nucleoside-Diphosphate Kinase; D015720:5'-Nucleotidase; D001081:Apyrase; C067869:CD39 antigen