Crystallization and preliminary diffraction analysis of a hyperthermostable DNA polymerase from a Thermococcus archaeon.
The hyperthermostable DNA polymerase from a marine Thermococcus archaeon has been crystallized in space group P212121, with unit-cell dimensions a = 94.8, b = 98.2, c = 112.2 A with one molecule per asymmetric unit. Conditions for data collection at 98 K have been identified, and a complete data set was collected to 2.2 A resolution. Strategies employed here may facilitate crystallization of other hyperthermostable proteins. The structure of this enzyme will provide the first structural data on the archaeal and hyperthermostable classes of DNA polymerases. Sequence homology to human polymerase alpha (polymerase B family) may make it a model for studying eukaryotic and viral polymerases and for the development of anti-cancer and anti-viral therapeutics.
Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
M
M
Zhou
C
C
Mao
A C
AC
Rodriguez
J R
JR
Kiefer
R B
RB
Kucera
L S
LS
Beese
D003460:Crystallization; D018360:Crystallography, X-Ray; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D019712:Thermococcus / Q000201:enzymology*
C070331:Tli polymerase; D004259:DNA-Directed DNA Polymerase