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. 1996 May;111(1):215–221. doi: 10.1104/pp.111.1.215

Purification and Characterization of a Cryoprotective Protein (Cryoprotectin) from the Leaves of Cold-Acclimated Cabbage.

F Sieg 1, W Schroder 1, J M Schmitt 1, D K Hincha 1
PMCID: PMC157828  PMID: 12226287

Abstract

We have purified a protein (cryoprotectin) from the leaves of cold-acclimated cabbage (Brassica oleracea L.) that protects thylakoids from nonacclimated spinach (Spinacia oleracea L.) against freeze-thaw damage. The procedure involves precipitations by heat, ammonium sulfate, and the glycosaminoglycan heparin and column chromatography on Polyamide 6 and a C18 reverse-phase matrix. After reverse-phase chromatography we obtained a single band of an apparent molecular mass of 7 kD when fractions that showed cryoprotective activity were analyzed by sodium dodecyl sulfate gel electrophoresis and silver staining. Gel-filtration experiments confirmed that the active protein is a monomer of 7 kD native molecular mass. This 7-kD protein could be purified only from cold-acclimated cabbage, but not from plants grown under nonacclimating conditions. Using peroxidase-labeled lectins, we show that cryoprotectin is a glycoprotein and that the saccharide moiety contains [alpha]1-3-linked fucose.

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Selected References

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  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Calvete J. J., Solís D., Sanz L., Díaz-Mauriño T., Töpfer-Petersen E. Glycosylated boar spermadhesin AWN-1 isoforms. Biological origin, structural characterization by lectin mapping, localization of O-glycosylation sites, and effect of glycosylation on ligand binding. Biol Chem Hoppe Seyler. 1994 Oct;375(10):667–673. doi: 10.1515/bchm3.1994.375.10.667. [DOI] [PubMed] [Google Scholar]
  3. Debray H., Decout D., Strecker G., Spik G., Montreuil J. Specificity of twelve lectins towards oligosaccharides and glycopeptides related to N-glycosylproteins. Eur J Biochem. 1981 Jun;117(1):41–55. doi: 10.1111/j.1432-1033.1981.tb06300.x. [DOI] [PubMed] [Google Scholar]
  4. Hincha D. K., Schmitt J. M. Long-term cryopreservation of thylakoid membranes. Methods Mol Biol. 1995;38:71–80. doi: 10.1385/0-89603-296-5:71. [DOI] [PubMed] [Google Scholar]
  5. Kohnke-Godt B., Gabius H. J. Heparin-binding lectin from human placenta: further characterization of ligand binding and structural properties and its relationship to histones and heparin-binding growth factors. Biochemistry. 1991 Jan 8;30(1):55–65. doi: 10.1021/bi00215a009. [DOI] [PubMed] [Google Scholar]
  6. Lynch D. V., Steponkus P. L. Plasma Membrane Lipid Alterations Associated with Cold Acclimation of Winter Rye Seedlings (Secale cereale L. cv Puma). Plant Physiol. 1987 Apr;83(4):761–767. doi: 10.1104/pp.83.4.761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Schägger H., von Jagow G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987 Nov 1;166(2):368–379. doi: 10.1016/0003-2697(87)90587-2. [DOI] [PubMed] [Google Scholar]
  8. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Volger H. G., Heber U. Cryoprotective leaf proteins. Biochim Biophys Acta. 1975 Dec 15;412(2):335–349. doi: 10.1016/0005-2795(75)90048-3. [DOI] [PubMed] [Google Scholar]

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