title: Purification, characterization, and crystallization of human pyrroline-5-carboxylate reductase.
abstract: Pyrroline-5-carboxylate reductase (P5CR) catalyzes the reduction of Delta1-pyrroline-5-carboxylate (P5C) to proline with concomitant oxidation of NAD(P)H to NAD(P)(+). The enzymatic cycle between P5C and proline is very important in many physiological and pathological processes. Human P5CR was over-expressed in Escherichia coli and purified to homogeneity by chromatography. Enzymatic assays of the wild-type protein were carried out using 3,4-dehydro-L-proline as substrate and NAD(+) as cofactor. The homopolymer was characterized by cross-linking and size exclusion gel filtration chromatography. Human P5CR was crystallized by the hanging-drop vapor-diffusion method at 37 degrees C. Diffraction data were obtained to a resolution of 2.8A and were suitable for high resolution X-ray structure determination.
authors.affiliation: Tsinghua-IBP Joint Research Group for Structural Biology, Tsinghua University, Beijing 100084, China.
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Meng
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Lou
authors.forename: Zhe
authors.initials: Z
authors.lastname: Liu
authors.forename: Dong
authors.initials: D
authors.lastname: Hui
authors.forename: Mark
authors.initials: M
authors.lastname: Bartlam
authors.forename: Zihe
authors.initials: Z
authors.lastname: Rao
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D011108:Polymers / Q000737:chemistry; D011392:Proline / Q000031:analogs & derivatives / Q000378:metabolism; D011762:Pyrroline Carboxylate Reductases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D012995:Solubility; D014867:Water
chemical_list: D011108:Polymers; D014867:Water; C002195:3,4-dehydroproline; D011392:Proline; D011762:Pyrroline Carboxylate Reductases