Purification, crystallization and data collection of the apoptotic nuclease endonuclease G

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Title
Purification, crystallization and data collection of the apoptotic nuclease endonuclease G
Author(s)
Sei Mee Yoon; Hyung Nam Song; Joon Hyuck Yang; M Y Lim; Y J Chung; Seong Eon Ryu; Eui-jeon Woo
Bibliographic Citation
Acta Crystallographica Section F-Structural Biology, vol. 65, no. 5, pp. 504-507
Publication Year
2009
Abstract
Endonuclease G (EndoG) is a mitochondrial enzyme that responds to apoptotic stimuli by translocating to the nucleus and cleaving chromosomal DNA. EndoG is the main apoptotic endonuclease in the caspase-independent pathway. Mouse EndoG without the mitochondrial localization signal (amino-acid residues 1-43) was successfully overexpressed, purified and crystallized using a microbatch method under oil. The initial crystal (type I) was grown in the presence of the detergent CTAB and diffracted to 2.8 A resolution, with unit-cell parameters a = 72.20, b = 81.88, c = 88.66 A, beta = 97.59 degrees in a monoclinic space group. The crystal contained two monomers in the asymmetric unit, with a predicted solvent content of 46.6%. Subsequent mutation of Cys110 improved the stability of the protein significantly and produced further crystals of types II, III and IV with space groups C2, P4(1)2(1)2 (or P4(3)2(1)2) and P2(1)2(1)2(1), respectively, in various conditions. This suggests the critical involvement of this conserved cysteine residue in the crystallization process.
Keyword
Endonuclease G
ISSN
1744-3091
Publisher
Int Union Crystallography
DOI
http://dx.doi.org/10.1107/S1744309109013335
Type
Article
Appears in Collections:
Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
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