Description
Chymotrypsin-like enzymes cleave their substrate proteins at the carboxy terminus of amino acids containing hydrophobic aliphatic or aromatic R-group side chains such as those found on leucine and phenylalanine amino acid structures [1]. The red phenylalanine FLISP probe, consisting of SR101-Phe-CMK, is simply a fluorescent-labeled analog of the early chymotrypsin-like enzyme inhibitor N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) [2].
Early studies using topoisomerase inhibitors in HL-60 cells indicated that TPCK, N-tosyl-L-lysine chloromethyl ketone (TLCK) and other serine protease inhibitors were able to prevent internucleosomal DNA degradation associated with apoptosis [3]. Dual staining experiments have used our green FAM-F-CMK (chymotrypsin detection probe) and red sulforhodamine (SR)-VAD-FMK FLICA caspase detection probe in the presence or absence of z-VAD-FMK, TPCK, or TLCK caspase or serine protease inhibitors [4-5]. This work demonstrated a significant inhibitory effect of the caspase inhibitor, z-VAD-FMK, on chymotrypsin-associated FAM-F-CMK binding in cells.
This group also showed that the chymotrypsin-like enzyme inhibitor, TPCK, had a strong suppressive effect on caspase activation and apoptosis induction. In contrast, the trypsin-like enzyme inhibitor, TLCK, had very little effect on caspase activation events as measured by the binding of the red FLICA probe SR-VAD-FMK in camptothecin-induced cell populations
Bioinformatics
| Alternate Names |
- FLISP SR101-Phe-CMK Serine Protease
- Phe-CMK Serine Protease
- TR-Phe-CMK (TRFCK)
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