TY - JOUR
T1 - Purification, crystallization and preliminary crystallographic analysis of a multiple cofactor-dependent DNA ligase from Sulfophobococcus zilligii
AU - Supangat, Supangat
AU - An, Young Jun
AU - Sun, Younguk
AU - Kwon, Suk Tae
AU - Cha, Sun Shin
PY - 2010/12
Y1 - 2010/12
N2 - A recombinant DNA ligase from Sulfophobococcus zilligii that shows multiple cofactor specificity (ATP, ADP and GTP) was expressed in Escherichia coli and purified under reducing conditions. Crystals were obtained by the microbatch crystallization method at 295 K in a drop containing 1 l protein solution (10 mg ml-1) and an equal volume of mother liquor [0.1 M HEPES pH 7.5, 10%(w/v) polyethylene glycol 10 000]. A data set was collected to 2.9 Å resolution using synchrotron radiation. The crystals belonged to space group P1, with unit-cell parameters a = 63.7, b = 77.1, c = 77.8 Å, = 83.4, Β = 82.4, = 74.6°. Assuming the presence of two molecules in the unit cell, the solvent content was estimated to be about 53.4%.
AB - A recombinant DNA ligase from Sulfophobococcus zilligii that shows multiple cofactor specificity (ATP, ADP and GTP) was expressed in Escherichia coli and purified under reducing conditions. Crystals were obtained by the microbatch crystallization method at 295 K in a drop containing 1 l protein solution (10 mg ml-1) and an equal volume of mother liquor [0.1 M HEPES pH 7.5, 10%(w/v) polyethylene glycol 10 000]. A data set was collected to 2.9 Å resolution using synchrotron radiation. The crystals belonged to space group P1, with unit-cell parameters a = 63.7, b = 77.1, c = 77.8 Å, = 83.4, Β = 82.4, = 74.6°. Assuming the presence of two molecules in the unit cell, the solvent content was estimated to be about 53.4%.
KW - DNA ligases
KW - Sulfophobococcus zilligii
KW - multiple cofactors
UR - http://www.scopus.com/inward/record.url?scp=78650123499&partnerID=8YFLogxK
U2 - 10.1107/S1744309110034135
DO - 10.1107/S1744309110034135
M3 - Article
C2 - 21139200
AN - SCOPUS:78650123499
SN - 1744-3091
VL - 66
SP - 1583
EP - 1585
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 12
ER -