Crystal structure of human pyrroline-5-carboxylate reductase.
Pyrroline-5-carboxylate reductase (P5CR) is a universal housekeeping enzyme that catalyzes the reduction of Delta(1)-pyrroline-5-carboxylate (P5C) to proline using NAD(P)H as the cofactor. The enzymatic cycle between P5C and proline is very important for the regulation of amino acid metabolism, intracellular redox potential, and apoptosis. Here, we present the 2.8 Angstroms resolution structure of the P5CR apo enzyme, its 3.1 Angstroms resolution ternary complex with NAD(P)H and substrate-analog. The refined structures demonstrate a decameric architecture with five homodimer subunits and ten catalytic sites arranged around a peripheral circular groove. Mutagenesis and kinetic studies reveal the pivotal roles of the dinucleotide-binding Rossmann motif and residue Glu221 in the human enzyme. Human P5CR is thermostable and the crystals were grown at 37 degrees C. The enzyme is implicated in oxidation of the anti-tumor drug thioproline.
Meng
Zhaohui
Z
Tsinghua-IBP Joint Research Group for Structural Biology, Tsinghua University, Beijing, China.
Lou
Zhiyong
Z
Liu
Zhe
Z
Li
Ming
M
Zhao
Xiaodong
X
Bartlam
Mark
M
Rao
Zihe
Z
D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D019281:Dimerization; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D017433:Protein Structure, Secondary; D011762:Pyrroline Carboxylate Reductases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D016415:Sequence Alignment; D013297:Streptococcus pyogenes / Q000201:enzymology; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase
D011762:Pyrroline Carboxylate Reductases; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase