title: zVAD-fmk, unlike BocD-fmk, does not inhibit caspase-6 acting on 14-3-3/Bad pathway in apoptosis of p815 mastocytoma cells.
abstract: In a preliminary study, we found that benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD- fmk), unlike Boc-aspartyl(OMe)-fluoromethylketone (BocD-fmk), at usual dosage could not prevent genistein-induced apoptosis of p815 mastocytoma cells. This study was undertaken to reveal the mechanism underlying the incapability of zVAD-fmk in preventing this type of apoptosis. We observed that 14-3-3 protein level was reduced in genistein-treated cells and that BocD-fmk but not zVAD-fmk prevented the reduction of 14-3-3 protein level and the release of Bad from 14-3-3. We also demonstrated that truncated Bad to Bcl-xL interaction in genistein- treated cells was prevented by BocD-fmk but not by zVAD-fmk treatment. Our data indicate that BocD- fmk, compared to zVAD-fmk, has a certain preference for inhibiting 14-3-3/Bad signalling pathway. We also elucidated that this differential efficacy of BocD-fmk and zVAD-fmk resulted from the different effect in inhibiting caspase-6 and that co-treatment of zVAD-fmk and caspase-6 specific inhibitor substantially prevented genistein-induced apoptosis. Our data shows that caspase-6 plays a role on Bad/14-3-3 pathway in genistein-induced apoptosis of p815 cells, and that the usual dose of zVAD-fmk, in contrast to BocD-fmk, did not prevent caspase-6 acting on 14-3-3/Bad-mediated event.
authors.affiliation: Department of Anatomy and Cell Biology (BK21 program), Dong-A University College of Medicine and Medical, Science Research Center, Busan 602-714, Korea.
authors.forename: Su-Bog
authors.initials: SB
authors.lastname: Yee
authors.forename: Soo Jin
authors.initials: SJ
authors.lastname: Baek
authors.forename: Hwan Tae
authors.initials: HT
authors.lastname: Park
authors.forename: Seung Hun
authors.initials: SH
authors.lastname: Jeong
authors.forename: Jin Hee
authors.initials: JH
authors.lastname: Jeong
authors.forename: Tae Hyun
authors.initials: TH
authors.lastname: Kim
authors.forename: Jong-Min
authors.initials: JM
authors.lastname: Kim
authors.forename: Byung Kap
authors.initials: BK
authors.lastname: Jeong
authors.forename: Bong Soo
authors.initials: BS
authors.lastname: Park
authors.forename: Taeg Kyu
authors.initials: TK
authors.lastname: Kwon
authors.forename: Il
authors.initials: I
authors.lastname: Yoon
authors.forename: Young Hyun
authors.initials: YH
authors.lastname: Yoo
mesh_terms: D048948:14-3-3 Proteins / Q000378:metabolism*; D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001593:Benzyl Compounds / Q000494:pharmacology*; D053178:Caspase 6 / Q000378:metabolism*; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D019833:Genistein / Q000494:pharmacology; D006845:Hydrocarbons, Fluorinated / Q000494:pharmacology*; D034801:Mastocytoma; D051379:Mice; D008928:Mitochondria / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D051025:bcl-Associated Death Protein / Q000378:metabolism*
chemical_list: D048948:14-3-3 Proteins; D000590:Amino Acid Chloromethyl Ketones; D001593:Benzyl Compounds; C476029:Boc-D-FMK; D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; D006845:Hydrocarbons, Fluorinated; D051025:bcl-Associated Death Protein; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D019833:Genistein; D053178:Caspase 6