Crystallization and preliminary X-ray crystallographic analysis of adenosine 5'-monophosphate deaminase (AMPD) from Arabidopsis thaliana in complex with coformycin 5'-phosphate.
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 P6(2)22, with unit-cell parameters a = b = 131.325, c = 208.254 A, alpha = beta = 90, gamma = 120 degrees. Diffraction data were collected to 3.34 A resolution from a crystal in complex with coformycin 5'-phosphate and to 4.05 A resolution from a crystal of a mercury derivative.
Han
Byung Woo
BW
Department of Biochemistry, University of Wisconsin-Madison, WI 53706-1544, USA.
Bingman
Craig A
CA
Mahnke
Donna K
DK
Sabina
Richard L
RL
Phillips
George N
GN
D000659:AMP Deaminase / Q000737:chemistry* / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry*; D003070:Coformycin / Q000737:chemistry / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D003625:Data Collection; D046911:Macromolecular Substances / Q000737:chemistry*; D010755:Organophosphates / Q000737:chemistry*
D029681:Arabidopsis Proteins; D046911:Macromolecular Substances; D010755:Organophosphates; D003070:Coformycin; D000659:AMP Deaminase