Caspase-dependant activation of chymotrypsin-like proteases mediates nuclear events during Jurkat T cell apoptosis
Section snippets
Methods and materials
Cell culture and treatments. Jurkat T cells were obtained from CATCC and maintained in RPMI 1640 medium (Sigma) supplemented with 10% heat-inactivated foetal calf serum, 2 mM glutamine, 100 U/ml penicillin and 100 mg/ml streptomycin in a humidified atmosphere of 5% CO2 in air at 37 °C. Cells were maintained in logarithmic growth phase by routine passage every 2–3 days. For induction of apoptosis, cells were seeded at a density of 106/ml in fresh cell culture medium and staurosporine was added to
Staurosporine-induced apoptosis in Jurkat T cells is accompanied by an increase in FFCK-binding proteins
Activation of non-caspase proteases has been implicated in a number of types of cell death including apoptosis, necrosis and autophagy [16], [39]. Therefore, in order to characterise the potential apoptotic roles of serine proteases, it is important to define the type of cell death in which they are active. In our initial studies we demonstrated that treatment of Jurkat T cells with 1 μM staurosporine (sts) for 2 h led to the development of features characteristic of apoptosis (Fig. 1A–C).
Acknowledgments
This work was funded by Science Foundation Ireland, Irish Council for Science and Engineering Technology, National Development Plan, Higher Education Authority, National University of Ireland, NUIG Millennium Research Fund, and Enterprise Ireland.
References (42)
History of the events leading to the formulation of the apoptosis concept
Toxicology
(2002)- et al.
A matter of life and death
Cancer Cell
(2002) - et al.
Membrane oligomerization and cleavage activates the caspase-8 (FLICE/MACHalpha1) death signal
J. Biol. Chem.
(1998) - et al.
Bcl-xL functions downstream of caspase-8 to inhibit Fas- and tumor necrosis factor receptor 1-induced apoptosis of MCF7 breast carcinoma cells
J. Biol. Chem.
(1998) - et al.
In the cut and thrust of apoptosis, serine proteases come of age
Biochem. Pharmacol.
(2003) - et al.
Granzyme B: pro-apoptotic, antiviral and antitumor functions
Curr. Opin. Immunol.
(2003) - et al.
Families of serine peptidases
Methods Enzymol.
(1994) - et al.
A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death
Mol. Cell
(2001) - et al.
The serine protease Omi/HtrA2 regulates apoptosis by binding XIAP through a reaper-like motif
J. Biol. Chem.
(2002) - et al.
HtrA2 promotes cell death through its serine protease activity and its ability to antagonize inhibitor of apoptosis proteins
J. Biol. Chem.
(2002)