Crystallization and preliminary x-ray crystallographic analysis of NAD+-dependent DNA ligase from Thermus filiformis.
A highly thermostable DNA ligase from Thermus filiformis has been crystallized at room temperature using methoxypolyethylene glycol 5000 as a precipitant. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 90.63, b = 117.80, c = 98. 65 A, beta = 115.56 degrees. Two molecules of DNA ligase are present in the asymmetric unit, giving a crystal volume per protein mass (V(m)) of 3.1 A(3) Da(-1) and a solvent content of 61%. A native data set extending to 3.0 A resolution has been collected at 100 K using synchrotron X-rays.
Lee
J Y
JY
Department of Chemistry, College of Natural Sciences, Division of Chemistry and Molecular Engineering, Seoul National University, Seoul 151--742, South Korea.
Kim
H K
HK
Chang
C
C
Eom
S H
SH
Hwang
K Y
KY
Cho
Y
Y
Yu
Y G
YG
Ryu
S E
SE
Kwon
S T
ST
Suh
S W
SW
D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D011088:DNA Ligases / Q000737:chemistry* / Q000302:isolation & purification; D004926:Escherichia coli; D008958:Models, Molecular; D009243:NAD / Q000737:chemistry; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D013824:Thermus / Q000201:enzymology*
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; D009243:NAD; D011088:DNA Ligases