Protein Expression and Purification 2006-09-01

Purification, characterization, and crystallization of human pyrroline-5-carboxylate reductase.

Zhaohui Meng, Zhiyong Lou, Zhe Liu, Dong Hui, Mark Bartlam, Zihe Rao

Index: Protein Expr. Purif. 49(1) , 83-7, (2006)

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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.

Related Compounds

  • L-3-Pyrroline-2-c...
  • 3,4-Dehydro-L-pro...

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