Purification, crystallization and preliminary crystallographic analysis of a Thermostable Endonuclease IV from Thermotoga maritima
The DNA repair enzyme Endonuclease IV from the thermophilic bacteriumThermotoga Maritima MSB8 (reference sequence: NC_000853) has been expressed in Escherichia coli and crystallized for X-ray analysis. Thermotoga maritima Endonuclease IV is a 287 amino-acid protein with 32% sequence identity to the Escherichia coli Endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting drop vapor-diffusion method. The protein crystallized in the space group P61, with a composition of one biological molecule in the asymmetric unit corresponding to a Mathew’s coefficient of 2.39 and a 47% solvent fraction. The unit-cell parameters for the crystals are a = 123.23 Å, b = 123.23 Å , c =35.34 Å, α = β= 90°, γ= 120°. Microseeding and further optimization yielded crystals with an X-ray diffraction limit of 2.4 Å. A single 70° data set was collected and processed resulting in an overall R merge and completeness of 9.5% and 99.3% respectively.
- Research Organization:
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-00OR22725
- OSTI ID:
- 1009913
- Journal Information:
- Acta Crystallographica. Section F, Vol. 65, Issue 12; ISSN 1744-3091
- Publisher:
- International Union of Crystallography
- Country of Publication:
- United States
- Language:
- English
Similar Records
Crystallization and preliminary crystallographic analysis of an esterase with a novel domain from the hyperthermophile Thermotoga maritima
Crystallization and preliminary X-ray crystallographic characterization of TrmFO, a folate-dependent tRNA methyltransferase from Thermotoga maritima