Purification, crystallization and preliminary X-ray crystallographic studies of the complex between Smc5 and the SUMO E3 ligase Mms21
Smc5/6, a protein complex that belongs to the structural maintenance of chromosome (SMC) family, plays a key role in DNA replication, sister chromatid recombination and DNA damage repair. The complex contains eight subunits, including a SUMO E3 ligase Mms21 (Nse2). The activity of Mms21 is important for regulation of Smc5/6 in the response to DNA damage. Mms21 and the Mms21-binding region of Smc5 were overexpressed and purified individually in Escherichia coli with a C-terminal LEHHHHHH tag. The Mms21-Smc5 protein complex was crystallized. The diffraction of the crystals was improved greatly by glutaraldehyde treatment. X-ray diffraction data sets were collected to resolutions of 2.3 and 3.9 {angstrom} from native and selenomethionine-derivative protein crystals, respectively. The crystals belonged to space group C2221, with unit-cell parameters a = 47.465, b = 97.574, c = 249.215 {angstrom} for the native crystals.
- Research Organization:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 1005872
- Journal Information:
- Acta Crystallogr. F, Vol. 65, Issue (8) ; 08, 2009; ISSN 1744-3091
- Country of Publication:
- United States
- Language:
- ENGLISH
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Related Subjects
99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE
CHROMATIDS
CHROMOSOMES
CRYSTALLIZATION
DIFFRACTION
DNA DAMAGES
DNA REPLICATION
ESCHERICHIA COLI
LIGASES
MAINTENANCE
PROTEINS
PURIFICATION
RECOMBINATION
REGULATIONS
REPAIR
SPACE GROUPS
X-RAY DIFFRACTION