Divalent-metal-dependent nucleolytic activity of Cu, Zn superoxide dismutase.
The known action of Cu, Zn superoxide dismutase (holo SOD) that converts O2- to O2 and H2O2 plays a crucial role in protecting cells from toxicity of oxidative stress. However, the overproduction of holo SOD does not result in increased protection but rather creates a variety of unfavorable effects, suggesting that too much holo SOD may be injurious to the cells. In the in vitro study, we report a finding that the holo SOD from bovine erythrocytes and its apo form possess a divalent-metal-dependent nucleolytic activity, which was confirmed by UV-vis absorption titration of calf thymus DNA (ctDNA) with the holo SOD, quenching of holo SOD intrinsic fluorescence by ctDNA, and by gel electrophoresis monitoring conversion of DNA from the supercoiled DNA to nicked and linear forms, and fragmentation of a linear lambdaDNA. Moreover, the DNA cleavage activity was examined in detail under certain reaction conditions. The steady-state study indicates that DNA cleavage supported by both forms of SOD obeys Michaelis-Menten kinetics. On the other hand, the assays with some other proteins indicate that this new function is specific to some proteins including the holo SOD. Therefore, this study reveals that the divalent-metal-dependent DNA cleavage activity is an intrinsic property of the holo SOD, which is independent of its natural metal (copper and zinc) sites, and may provide an alternative insight into the link between SOD enzymes and neurodegenerative disorders.
Jiang
Wei
W
Department of Chemistry, Huazhong University of Science and Technology, Wuhan, 430074, China.
Shen
Tao
T
Han
Yingchun
Y
Pan
Qunhui
Q
Liu
Changlin
C
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000187:drug effects*; D003300:Copper / Q000737:chemistry*; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology*; D016166:Free Radical Scavengers / Q000737:chemistry; D006801:Humans; D007700:Kinetics; D011489:Protein Denaturation; D013482:Superoxide Dismutase / Q000737:chemistry* / Q000494:pharmacology / Q000633:toxicity; D015032:Zinc / Q000737:chemistry*
D004268:DNA-Binding Proteins; D016166:Free Radical Scavengers; D003300:Copper; D004247:DNA; C427203:calf thymus DNA; D013482:Superoxide Dismutase; D015032:Zinc