Expression of mutated cationic trypsinogen reduces cellular viability in AR4-2J cells.
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.
Gaiser
Sebastian
S
Medizinische Klinik und Poliklinik II, Universitätsklinikum Leipzig AöR, Ph.-Rosenthal-Str. 27, 04103 Leipzig, Germany.
Ahler
Astrid
A
Gundling
Felix
F
Kruse
Marie-Luise
ML
Savkovic
Vuk
V
Selig
Lena
L
Teich
Niels
N
Tomasini
Richard
R
Dagorn
Jean-Charles
JC
Mössner
Joachim
J
Keim
Volker
V
Bödeker
Hans
H
D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D002470:Cell Survival; D016297:Mutagenesis, Site-Directed; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism*; D014362:Trypsinogen / Q000378:metabolism*
D011994:Recombinant Proteins; D014362:Trypsinogen; C489303:PRSS1 protein, human; D014357:Trypsin