TY - JOUR
T1 - Purification, crystallization and preliminary crystallographic analysis of a thermostable endonuclease IV from Thermotoga maritima
AU - Hughes, Ronny C.
AU - Tomanicek, Stephen J.
AU - Ng, Joseph D.
AU - Coates, Leighton
PY - 2009
Y1 - 2009
N2 - The DNA-repair enzyme endonuclease IV from the thermophilic bacterium Thermotoga maritima MSB8 (reference sequence NC-000853) has been expressed in Escherichia coli and crystallized for X-ray analysis. T. maritima endonuclease IV is a 287-amino-acid protein with 32% sequence identity to E. coli endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting-drop vapor-diffusion method. The protein crystallized in space group P61, with one biological molecule in the asymmetric unit, corresponding to a Matthews coefficient of 2.39 Å3 Da-1 and 47% solvent content. The unit-cell parameters of the crystals were a = b = 123.2, c = 35.6 Å. Microseeding and further optimization yielded crystals with an X-ray diffraction limit of 2.36 Å. A single 70° data set was collected and processed, resulting in an overall Rmerge and a completeness of 9.5% and 99.3%, respectively.
AB - The DNA-repair enzyme endonuclease IV from the thermophilic bacterium Thermotoga maritima MSB8 (reference sequence NC-000853) has been expressed in Escherichia coli and crystallized for X-ray analysis. T. maritima endonuclease IV is a 287-amino-acid protein with 32% sequence identity to E. coli endonuclease IV. The protein was purified to homogeneity and was crystallized using the sitting-drop vapor-diffusion method. The protein crystallized in space group P61, with one biological molecule in the asymmetric unit, corresponding to a Matthews coefficient of 2.39 Å3 Da-1 and 47% solvent content. The unit-cell parameters of the crystals were a = b = 123.2, c = 35.6 Å. Microseeding and further optimization yielded crystals with an X-ray diffraction limit of 2.36 Å. A single 70° data set was collected and processed, resulting in an overall Rmerge and a completeness of 9.5% and 99.3%, respectively.
KW - DNA-repair proteins
KW - Endonuclease IV
KW - Thermotoga maritima
UR - http://www.scopus.com/inward/record.url?scp=74549223044&partnerID=8YFLogxK
U2 - 10.1107/S1744309109047393
DO - 10.1107/S1744309109047393
M3 - Article
C2 - 20054139
AN - SCOPUS:74549223044
SN - 1744-3091
VL - 65
SP - 1317
EP - 1319
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 12
ER -