Expression of mutated cationic trypsinogen reduces cellular viability in AR4-2J cells |
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Authors: | Gaiser Sebastian Ahler Astrid Gundling Felix Kruse Marie-Luise Savkovic Vuk Selig Lena Teich Niels Tomasini Richard Dagorn Jean-Charles Mössner Joachim Keim Volker Bödeker Hans |
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Affiliation: | Medizinische Klinik und Poliklinik II, Universit?tsklinikum Leipzig A?R, Ph.-Rosenthal-Str. 27, 04103 Leipzig, Germany. |
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Abstract: | Mutations in the human cationic trypsinogen are associated with hereditary pancreatitis. The cDNA coding for human cationic trypsinogen was subcloned into the expression vector pcDNA3. The mutations R122H, N29I, A16V, D22G, and K23R were introduced by site directed mutagenesis. We constructed an expression vector coding for active trypsin by subcloning the cDNA of trypsin lacking the coding region for the trypsin activating peptide behind an appropriate signal peptide. Expression of protein was verified by Western blot and measurement of enzymatic activity. AR4-2J cells were transiently transfected with the different expression vectors and cell viability and intracellular caspase-3 activity were quantified. In contrast to wild-type trypsinogen, expression of active trypsin and mutated trypsinogens reduced cell viability of AR4-2J cells. Expression of trypsin and R122H trypsinogen induced caspase-3 activity. Acinar cells might react to intracellular trypsin activity by triggering apoptosis. |
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Keywords: | Hereditary pancreatitis R122H trypsinogen Trypsin Apoptosis Caspase-3 |
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