Open AccessCrystallization and X‐ray diffraction analysis of the RNA primer/promoter‐binding domain of influenza A virus RNA‐dependent RNA polymerase PB2
Author(s) -
Kuzuhara Takashi,
Kise Daisuke,
Yoshida Hiroko,
Horita Takahiro,
Murazaki Yoshimi,
Utsunomiya Hiroko,
Tsuge Hideaki
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108042942
Subject(s) - primer (cosmetics) , rna , rna dependent rna polymerase , rna polymerase i , polymerase , rna polymerase , crystallization , microbiology and biotechnology , virology , crystallography , chemistry , biology , dna , gene , genetics , organic chemistry
The C‐terminal domain protein (amino‐acid residues 535–759) of the PB2 subunit of the RNA‐dependent RNA polymerase from the highly pathogenic influenza A virus was expressed as a soluble protein in Escherichia coli and crystallized using sodium formate as a precipitant. Data sets were collected from crystals of native and selenomethionine‐substituted protein on the KEK NW12 beamline at the Photon Factory and the crystals diffracted to a maximum resolution of 2.44 Å for the SeMet‐derivative crystal. The native crystals were found to belong to space group P 3 2 21, with unit‐cell parameters a = b = 52.5, c = 156.3 Å. The Matthews value ( V M ) was 2.7 Å 3 Da −1 , assuming the presence of one molecule in the asymmetric unit. The SeMet‐derivative crystals were found to belong to the same space group, with unit‐cell parameters a = b = 52.6, c = 156.4 Å. Attempts are being made to solve the structure by multi‐wavelength anomalous dispersion phasing.