Enzymatic active site of caspase-activated DNase (CAD) and its inhibition by inhibitor of CAD.
作者： Hideki Sakahira , Yasutaka Takemura , Shigekazu Nagata
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摘要: Abstract Caspase-activated DNase (CAD) is a deoxyribonuclease that causes DNA fragmentation during apoptosis. In proliferating cells, CAD complexed with ICAD (inhibitor of CAD) and its activity suppressed. Here, we established quantitative assay for measures the number 3′ hydroxyl groups on CAD-generated fragments. Chemical modification histidine residues substrate protection experiments demonstrated presence reactive within active site enzyme. Analysis by site-directed mutagenesis suggested at least four in C-terminal part molecule are essential catalytic DNase. did not protect from chemical residues, indicating it does mask CAD. contrast, blocked ability to bind DNA, suggesting steric or electrostatic hindrance DNA. This molecular mechanism inhibition similar proposed colicin endonuclease inhibitor, immunity protein.
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Edith Wilson Miles, Modification of histidyl residues in proteins by diethylpyrocarbonate. Methods in Enzymology. ,vol. 47, pp. 431- 442 ,(1977) , 10.1016/0076-6879(77)47043-5
Hideki Sakahira, Masato Enari, Shigekazu Nagata, Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis Nature. ,vol. 391, pp. 96- 99 ,(1998) , 10.1038/34214
D. Suck, C. Oefner, W. Kabsch, Three-dimensional structure of bovine pancreatic DNase I at 2.5 A resolution. The EMBO Journal. ,vol. 3, pp. 2423- 2430 ,(1984) , 10.1002/J.1460-2075.1984.TB02149.X
Fernando Rodrigues-Lima, Amanda C. Fensome, Michelle Josephs, Joe Evans, Robert J. Veldman, Matilda Katan, Structural Requirements for Catalysis and Membrane Targeting of Mammalian Enzymes with Neutral Sphingomyelinase and Lysophospholipid Phospholipase C Activities ANALYSIS BY CHEMICAL MODIFICATION AND SITE-DIRECTED MUTAGENESIS Journal of Biological Chemistry. ,vol. 275, pp. 28316- 28325 ,(2000) , 10.1074/JBC.M003080200
Takanori Otomo, Hideki Sakahira, Koichi Uegaki, Shigekazu Nagata, Toshio Yamazaki, Structure of the heterodimeric complex between CAD domains of CAD and ICAD. Nature Structural & Molecular Biology. ,vol. 7, pp. 658- 662 ,(2000) , 10.1038/77957
Colin Kleanthous, Andrew M. Hemmings, Ulrike C. Kühlmann, Ansgar J. Pommer, Neil Ferguson, Sheena E. Radford, Geoffrey R. Moore, Richard James, Structural and mechanistic basis of immunity toward endonuclease colicins Nature Structural & Molecular Biology. ,vol. 6, pp. 243- 252 ,(1999) , 10.1038/6683
Masato Enari, Hideki Sakahira, Hideki Yokoyama, Katsuya Okawa, Akihiro Iwamatsu, Shigekazu Nagata, A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD Nature. ,vol. 391, pp. 43- 50 ,(1998) , 10.1038/34112
Armin Lahm, Dietrich Suck, DNase I-induced DNA conformation: 2 Å Structure of a DNase I-octamer complex Journal of Molecular Biology. ,vol. 222, pp. 645- 667 ,(1991) , 10.1016/0022-2836(91)90502-W
John S. McCarty, Shen Yon Toh, Peng Li, Multiple domains of DFF45 bind synergistically to DFF40: roles of caspase cleavage and sequestration of activator domain of DFF40. Biochemical and Biophysical Research Communications. ,vol. 264, pp. 181- 185 ,(1999) , 10.1006/BBRC.1999.1498
Shigekazu Nagata, APOPTOTIC DNA FRAGMENTATION Experimental Cell Research. ,vol. 256, pp. 12- 18 ,(2000) , 10.1006/EXCR.2000.4834