拟南芥5'-单磷酸腺苷脱氨酶（AMPD）与5'-磷酸助间型霉素复合物的结晶及初步X射线晶体学分析
Crystallization and preliminary X-ray crystallographic analysis of adenosine 5'-monophosphate deaminase (AMPD) from Arabidopsis thaliana in complex with coformycin 5'-phosphate.
作者信息
Han Byung Woo, Bingman Craig A, Mahnke Donna K, Sabina Richard L, Phillips George N
机构信息
Department of Biochemistry, University of Wisconsin-Madison, WI 53706-1544, USA.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2005 Aug 1;61(Pt 8):740-2. doi: 10.1107/S1744309105019792. Epub 2005 Jul 8.
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.
5'-单磷酸腺苷脱氨酶（AMPD）是一种真核酶，可将5'-单磷酸腺苷（AMP）转化为5'-肌苷酸（IMP）和氨。将来自拟南芥的AMPD（AtAMPD）克隆到杆状病毒转移载体p2Bac中，并与修饰的杆状病毒基因组一起共转染到草地贪夜蛾（Sf9）细胞中。所得重组杆状病毒经噬菌斑纯化、扩增后用于过表达重组AtAMPD。已获得纯化的AtAMPD晶体，其结合了过渡态抑制剂5'-磷酸助间型霉素。晶体属于空间群P6(2)22，晶胞参数a = b = 131.325，c = 208.254 Å，α = β = 90°，γ = 120°。从与5'-磷酸助间型霉素复合物的晶体收集到3.34 Å分辨率的衍射数据，从汞衍生物晶体收集到4.05 Å分辨率的衍射数据。
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