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. 1986 Jan;80(1):142–144. doi: 10.1104/pp.80.1.142

Purification to Homogeneity of Pyrroline-5-Carboxylate Reductase of Barley

Rolf Krueger 1,2, Hans-Jürgen Jäger 1,2, Martin Hintz 1,2, Edwin Pahlich 1,2
PMCID: PMC1075072  PMID: 16664571

Abstract

An enzyme has been purified to homogeneity from barley seedlings which has `proline dehydrogenase' and the pyrroline-5-carboxylic acid reductase activities. The purification achieved is 39,000-fold as calculated from the proline dehydrogenase activity. The subunit molecular weight of the protein is 30 kilodaltons. The native enzyme has molecular weights up to 480 kilodaltons, depending on the buffer environment. From the pH profiles, the specific activities and thermodynamic considerations, it is concluded that the plant proline dehydrogenase functions in vivo as a pyrroline-5-carboxylate reductase.

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Selected References

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  1. ADAMS E., GOLDSTONE A. Hydroxyproline metabolism. III. Enzymatic synthesis of hydroxyproline from Delta 1-pyrroline-3-hydroxy-5-carboxylate. J Biol Chem. 1960 Dec;235:3499–3503. [PubMed] [Google Scholar]
  2. Costilow R. N., Cooper D. Identity of proline dehydrogenase and delta1-pyrroline-5-carboxylic acid reductase in Clostridium sporogenes. J Bacteriol. 1978 Apr;134(1):139–146. doi: 10.1128/jb.134.1.139-146.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  4. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  5. MEISTER A., RADHAKRISHNAN A. N., BUCKLEY S. D. Enzymatic synthesis of L-pipecolic acid and L-proline. J Biol Chem. 1957 Dec;229(2):789–800. [PubMed] [Google Scholar]
  6. Matsuzawa T., Ishiguro I. Delta 1-pyrroline-5-carboxylate reductase from baker's yeast: further purification by affinity chromatography with 5' AMP-Sepharose 4B. Biochim Biophys Acta. 1980 Dec 4;616(2):381–383. doi: 10.1016/0005-2744(80)90155-2. [DOI] [PubMed] [Google Scholar]
  7. Matsuzawa T., Ishiguro I. delta 1-Pyrroline-5-carboxylate reductase from Baker's yeast. Purification, properties and its application in the assays of L-delta 1-pyrroline-5-carboxylate and L-ornithine in tissue. Biochim Biophys Acta. 1980 Jun 13;613(2):318–323. doi: 10.1016/0005-2744(80)90086-8. [DOI] [PubMed] [Google Scholar]
  8. Meile L., Leisinger T. Purification and properties of the bifunctional proline dehydrogenase/1-pyrroline-5-carboxylate dehydrogenase from Pseudomonas aeruginosa. Eur J Biochem. 1982 Dec;129(1):67–75. doi: 10.1111/j.1432-1033.1982.tb07021.x. [DOI] [PubMed] [Google Scholar]
  9. Nielsen B. L., Brown L. R. The basis for colored silver-protein complex formation in stained polyacrylamide gels. Anal Biochem. 1984 Sep;141(2):311–315. doi: 10.1016/0003-2697(84)90047-2. [DOI] [PubMed] [Google Scholar]
  10. PEISACH J., STRECKER H. J. The interconversion of glutamic acid and proline. V. The reduction of delta 1-pyrroline-5-carboxylic acid to proline. J Biol Chem. 1962 Jul;237:2255–2260. [PubMed] [Google Scholar]
  11. SMITH M. E., GREENBERG D. M. Characterization of an enzyme reducing pyrroline-5-carboxylate to proline. Nature. 1956 Jun 16;177(4520):1130–1130. doi: 10.1038/1771130a0. [DOI] [PubMed] [Google Scholar]
  12. SMITH M. E., GREENBERG D. M. Preparation and properties of partially purified glutamic semialdehyde reductase. J Biol Chem. 1957 May;226(1):317–327. [PubMed] [Google Scholar]
  13. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  14. Williams I., Frank L. Improved chemical synthesis and enzymatic assay of delta-1-pyrroline-5-carboxylic acid. Anal Biochem. 1975 Mar;64(1):85–97. doi: 10.1016/0003-2697(75)90408-x. [DOI] [PubMed] [Google Scholar]
  15. YURA T., VOGEL H. J. Pyrroline-5-carboxylate reductase of Neurospora crassa; partial purification and some properties. J Biol Chem. 1959 Feb;234(2):335–338. [PubMed] [Google Scholar]

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