Open AccessCrystallization and preliminary X‐ray crystallographic analysis of adenosine 5′‐monophosphate deaminase (AMPD) from Arabidopsis thaliana in complex with coformycin 5′‐phosphate
Author(s) -
Han Byung Woo,
Bingman Craig A.,
Mahnke Donna K.,
Sabina Richard L.,
Phillips George N.
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105019792
Subject(s) - adenosine monophosphate , sf9 , microbiology and biotechnology , recombinant dna , biology , adenosine , chemistry , biochemistry , spodoptera , gene
Adenosine 5′‐monophosphate deaminase (AMPD) is a eukaryotic enzyme that converts adenosine 5′‐monophosphate (AMP) to inosine 5′‐monophosphate (IMP) and ammonia. AMPD from Arabidopsis thaliana (AtAMPD) was cloned into the baculoviral transfer vector p2Bac and co‐transfected along with a modified baculoviral genome into Spodoptera frugiperda (Sf9) cells. The resulting recombinant baculovirus were plaque‐purified, amplified and used to overexpress recombinant AtAMPD. Crystals of purified AtAMPD have been obtained to which coformycin 5′‐phosphate, a transition‐state inhibitor, is bound. Crystals belong to space group P 6 2 22, with unit‐cell parameters a = b = 131.325, c = 208.254 Å, α = β = 90, γ = 120°. Diffraction data were collected to 3.34 Å resolution from a crystal in complex with coformycin 5′‐phosphate and to 4.05 Å resolution from a crystal of a mercury derivative.