TY - JOUR
T1 - Purification, crystallization and initial crystallographic analysis of RNA polymerase holoenzyme from Thermus thermophilus
AU - Vassylyeva, Marina N.
AU - Lee, Jookyung
AU - Sekine, Shun Ichi
AU - Laptenko, Oleg
AU - Kuramitsu, Seiki
AU - Shibata, Takehiko
AU - Inoue, Yorinao
AU - Borukhov, Sergei
AU - Vassylyev, Dmitry G.
AU - Yokoyama, Shigeyuki
PY - 2002/9
Y1 - 2002/9
N2 - RNA polymerase holoenzyme from Thermus thermophilus, consisting of six protein subunits (α2, β, β′, ω and σ70) and having a total molecular mass of about 450 kDa, was purified and crystallized by the hanging-drop vapour-diffusion technique under mild near-physiological conditions. The crystals diffract beyond 3 Å resolution. Careful analysis of diffraction data revealed that the crystals belong to space group P32, with unit-cell parameters a = b = 236.35, c = 249.04 Å, and have perfect twinning along the threefold axis. A complete data set at 3 Å resolution was collected and an unambiguous molecular-replacement solution was found using the structure of T. aquaticus RNA polymerase core enzyme as a search model. The refinement of structure and model building of the σ70 subunit is now in progress.
AB - RNA polymerase holoenzyme from Thermus thermophilus, consisting of six protein subunits (α2, β, β′, ω and σ70) and having a total molecular mass of about 450 kDa, was purified and crystallized by the hanging-drop vapour-diffusion technique under mild near-physiological conditions. The crystals diffract beyond 3 Å resolution. Careful analysis of diffraction data revealed that the crystals belong to space group P32, with unit-cell parameters a = b = 236.35, c = 249.04 Å, and have perfect twinning along the threefold axis. A complete data set at 3 Å resolution was collected and an unambiguous molecular-replacement solution was found using the structure of T. aquaticus RNA polymerase core enzyme as a search model. The refinement of structure and model building of the σ70 subunit is now in progress.
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U2 - 10.1107/S0907444902011770
DO - 10.1107/S0907444902011770
M3 - Article
C2 - 12198314
AN - SCOPUS:14244270520
SN - 0907-4449
VL - 58
SP - 1497
EP - 1500
JO - Acta Crystallographica Section D: Biological Crystallography
JF - Acta Crystallographica Section D: Biological Crystallography
IS - 9
ER -