A Novel Apoptotic Protease Activated in Human Breast Cancer Cells after Poisoning Topoisomerase I
Annual rept. 15 Sep 1999-14 Sep 2000
CASE WESTERN RESERVE UNIV CLEVELAND OH
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The goal of this grant is to clone the unknown protease activated by the active anti-breast cancer agent, Beta-lapachone Beta-lap. The research team showed for the first time that Beta-lap requires NQ01 1, a two-electron reduction enzyme elevated in many human breast cancers, for activation. The team then characterized the unknown apoptotic protease activated in human breast cancer cells by Beta-lap, defining endpoints which will be essential for the ultimate isolation of this novel apoptotic protease. The unknown protease a is a non-caspase cytsteine protease b cleaves p53, lamin Beta and PARP atypically in an NQO1-dependent manner at a time co-incident with calpain activation appearance of an 18 kDa active form and its movement into the nucleus by confocal microscopy and c is calcium-dependent, where BAPTA-AM and EDTA blocked cell death caused by Beta-lap. Cloning the unknown protease activated by Beta- lap is ongoing by this research team using standard biochemical methodology and p53 andor PARP cleavage site identification.
- Genetic Engineering and Molecular Biology
- Anatomy and Physiology
- Medicine and Medical Research