Apoptosis of carrot nuclei inin vitro system induced by cytochrome c

The apoptosis of carrot nuclei induced in cytosol of carrot cells by cytochrome c is reported. This kind of cell-free apoptosis system of plant, CS-100, is an efficient In vitro inducing system. Several typical characteristics of apoptosis including chromatin condensation, margina-tion and apoptotic bodies could be detected, and the degradation of genome in carrot nuclei into nucleosomal fragments could be detected. TUNEL assay shows the specific breakage of 3'-OH ends of DNA strand. The results indicate that CS-100 is an efficient and easily-operated in vitro system to study the apoptotic mechanism in plant and the apoptosis in plant cells may share similar pathways to apoptosis in animal cells.     

猪肝胆绿素还原酶的纯化及某些性质的研究

通过高速离心(105,000×g),硫酸铵分级分离、NADP—agaxose层析和SephadexG—100层析,从猪肝的胞浆中分离获得胆绿素还原酶.经SDS—聚丙烯酰胺凝胶电泳表明,该酶达到均一程度,分子量约为47,000道尔顿.酶被纯化4200倍,回收率为38%.这种酶利用NADPH和NADH作为电子供体,对巯基试剂特别敏感.如DTNB、NEM、pCMB及IDA等,都能不同程度地抑制该酶的活性.由HgCl_2处理,该酶活性的抑制是不可逆的.该酶用NADPH或NADH作电子供体时的最适pH分别是pH7.4和pH7.0.     

Induction of Apoptosis in Purified Nuclei from Tobacco-Suspension Cells by Cytochrome b6／f Complex

An apoptotic cell-free system containing cytosol and nuclei from normally cultured tobacco suspension cells was used to show that a spinach chloroplast preparation can induce apoptosis in nuclei, evidenced by DNA electrophoresis and fluorescence microscopy observations, Further study showed that the chloroplast preparation or its pellet (thylakoid membrane) after hypoosmotic or supersonic treatment still exhibited the apoptosis-inducing activity, but the supernatant had no effect, which indicates that the apoptosisinducing effector in the chloroplast preparation is water-insoluble. The induction of apoptosis by chloroplast preparation could be attenuated by Ac-DEVD-CHO, the specific inhibitor of Caspase-3, implying involvement of a Caspase-3-1ike protease during the process. Furthermore, extensive apoptosis in nuclei was induced by cytochrome b6/f on the thylakoid membrane, indicating that this important cytochrome complex may have an important role in the chloroplast-related apoptotic pathway.     

Flavonoids as inhibitors of rat liver cytosolic glutathione S-transferase

Summary The inhibitory potencies of different flavonoids for rat liver cytosolic glutathione S-transferase activity varied over 30-fold, depending on the pattern of hydroxylation, the presence of a C-2, C-3 double bond and the substitution of a hydroxyl group with a sugar moiety. Kinetic inactivation studies of the enzyme with the inhibitor quercetin revealed a non-competitive profile versus both glutathione and 1-chloro-2,4-dinitrobenzene.